Biphenyl derivatives

ABSTRACT

This invention provides biphenyl derivatives of formula I:  
                 
         wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , W, a, b and c are as defined in the specification, or a pharmaceutically acceptable salt or solvate or stereoisomer thereof. The biphenyl derivatives of this invention possess both β 2  adrenergic receptor agonist and muscarinic receptor antagonist activity and therefore, such biphenyl derivatives are useful for treating pulmonary disorders, such as chronic obstructive pulmonary disease and asthma.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.60/447,843, filed on Feb. 14, 2003; and U.S. Provisional Application No.60/467,035, filed on May 1, 2003; the entire disclosures of which areincorporated herein by reference in their entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to novel biphenyl derivatives that areuseful for treating pulmonary disorders. This invention also relates topharmaceutical compositions comprising such biphenyl derivatives,processes and intermediates for preparing such biphenyl derivatives andmethods of using such biphenyl derivatives to treat pulmonary disorders.

2. State of the Art

Pulmonary disorders, such as asthma and chronic obstructive pulmonarydisease (COPD), are commonly treated with bronchodilators. One class ofbronchodilator in widespread use consists of β₂ adrenergic receptor(adrenoceptor) agonists, such as albuterol, formoterol and salmeterol.These compounds are generally administered by inhalation. Another classof bronchodilator consists of muscarinic receptor antagonists(anticholinergic compounds), such as ipratropium and tiotropium. Thesecompounds are also typically administered by inhalation.

Pharmaceutical compositions containing both a β₂ adrenergic receptoragonist and a muscarinic receptor antagonist are also known in the artfor use in treating pulmonary disorders. For example, U.S. Pat. No.6,433,027 discloses medicament compositions containing a muscarinicreceptor antagonist, such as tiotropium bromide, and a β₂ adrenergicreceptor agonist, such as formoterol fumarate.

Although compounds having either β₂ adrenergic receptor agonist ormuscarinic receptor antagonist activity are known, no compound havingboth β₂ adrenergic receptor agonist and muscarinic receptor antagonistactivity has been previously disclosed. Compounds possessing both β₂adrenergic receptor agonist and muscarinic receptor antagonist activityare highly desirable since such bifunctional compounds would providebronchodilation through two independent modes of action while havingsingle molecule pharmacokinetics.

SUMMARY OF THE INVENTION

The present invention provides novel biphenyl derivatives that areuseful for treating pulmonary disorders. Among other properties,compounds of this invention have been found to possess both β₂adrenergic receptor agonist and muscarinic receptor antagonist activity.

Accordingly, in one of its composition aspects, the present invention isdirected to a compound of formula I:

wherein:

a is 0 or an integer of from 1 to 3;

each R¹ is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(1a), —C(O)OR^(1b),—SR^(1c), —S(O)R^(1d), S(O)₂R^(1e) and —NR^(1f)R^(1g);

each of R^(1a), R^(1b), R^(1c), R^(1d), R^(1e), R^(1f) and R^(1g) isindependently hydrogen, (1-4C)alkyl or phenyl-(1-4C)alkyl;

b is 0 or an integer of from 1 to 3;

each R² is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(2a), —C(O)OR^(2b),—SR^(2c), —S(O)R^(2d), S(O)₂R^(2e) and —NR^(2f)R^(2g);

each of R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g) isindependently hydrogen, (1-4C)alkyl or phenyl-(1-4C)alkyl;

W is attached to the 3- or 4-position with respect to the nitrogen atomin the piperidine ring and represents O or NW^(a);

W^(a) is hydrogen or (1-4C)alkyl;

c is 0 or an integer of from 1 to 4;

each R³ is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(3a), —C(O)OR^(3b),—SR^(3c), —S(O)R^(3d), S(O)₂R^(3e) and —NR^(3f)R^(3g); or two R³ groupsare joined to form (1-3C)alkylene, (2-3C)alkenylene or oxiran-2,3-diyl;

each of R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3f) and R^(3g) isindependently hydrogen or (1-4C)alkyl;

R⁴ is a divalent group of the formula:—(R^(4a))_(d)-(A¹)_(e)-(R^(4b))_(f)-Q-(R^(4c))_(g)-(A²)_(h)-(R^(4d))_(i)—wherein

d, e, f, g, h and i are each independently selected from 0 and 1;

R^(4a), R^(4b), R^(4c) and R^(4d) are each independently selected from(1-10C)alkylene, (2-10C)alkenylene and (2-10C)alkynylene, wherein eachalkylene, alkenylene or alkynylene group is unsubstituted or substitutedwith from 1 to 5 substituents independently selected from (1-4C)alkyl,fluoro, hydroxy, phenyl and phenyl-(1-4C)alkyl;

A¹ and A² are each independently selected from (3-7C)cycloalkylene,(6-10C)arylene, —O-(6-10C)arylene, (6-10C)arylene-O—,(2-9C)heteroarylene, —O-(2-9C)heteroarylene, (2-9C)heteroarylene-O— and(3-6C)heterocyclene, wherein each cycloalkylene is unsubstituted orsubstituted with from 1 to 4 substitutents selected independently from(1-4C)alkyl, and each arylene, heteroarylene or heterocyclene group isunsubstituted or substituted with from 1 to 4 substituents independentlyselected from halo, (1-4C)alkyl, (1-4C)alkoxy, —S-(1-4C)alkyl,—S(O)—(1-4C)alkyl, —S(O)₂-(1-4C)alkyl, —C(O)O(1-4C)alkyl, carboxy,cyano, hydroxy, nitro, trifluoromethyl and trifluoromethoxy;

Q is selected from a bond, —O—, —C(O)O—, —OC(O)—, —S—, —S(O)—, —S(O)₂—,—N(Q^(a))C(O), —C(O)N(Q^(b))-, —N(Q^(c))S(O)₂—, —S(O)₂N(Q^(d))-,—N(Q^(f))C(O)N(Q^(f))-, —N(Q^(g))S(O)₂N(Q^(h))-, —OC(O)N(Q^(i))-,—N(Q^(j))C(O)O— and —N(Q^(k));

Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g), Q^(h), Q^(i), Q^(j) andQ^(k) are each independently selected from hydrogen, (1-6C)alkyl, A³ and(1-4C)alkylene-A⁴, wherein the alkyl group is unsubstituted orsubstituted with from 1 to 3 substituents independently selected fromfluoro, hydroxy and (1-4C)alkoxy; or together with the nitrogen atom andthe group R^(4b) or R^(4c) to which they are attached, form a 4-6membered azacycloalkylene group;

A³ and A⁴ are each independently selected from (3-6C)cycloalkyl,(6-10C)aryl, (2-9C)heteroaryl and (3-6C)heterocyclyl, wherein eachcycloalkyl is unsubstituted or substituted with from 1 to 4substitutents selected independently from (1-4C)alkyl and each aryl,heteroaryl or heterocyclyl group is unsubstituted or substituted withfrom 1 to 4 substituents independently selected from halo, (1-4C)alkyland (1-4C)alkoxy;

provided that the number of contiguous atoms in the shortest chainbetween the two nitrogen atoms to which R⁴ is attached is in the rangeof from 4 to 16;

R⁵ represents hydrogen or (1-4C)alkyl;

R⁶ is —NR^(6a)CR^(6b)(O) or —CR^(6c)R^(6d)OR^(6e) and R⁷ is hydrogen; orR⁶ and R⁷ together form —NR^(7a)C(O)—CR^(7b)═CR^(7c)—,—CR^(7d)═CR^(7e)—C(O)—NR^(7f)—,—NR^(7g)C(O)—CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p)—;

each of R^(6a), R^(6b), R^(6c), R^(6d) and R^(6e) is independentlyhydrogen or (1-4C)alkyl; and

each of R^(7a), R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(7g), R^(7h),R^(7i), R^(7j), R^(7k), R^(7l), R^(7m), R^(7n), R^(7o) and R^(7p) isindependently hydrogen or (1-4C)alkyl;

or a pharmaceutically acceptable salt or solvate or stereoisomerthereof.

In another of its composition aspects, this invention is directed to acompound of formula II:

wherein

R⁴ is as defined herein (including any specific or preferredembodiments);

W represents O or NH;

or a pharmaceutically acceptable salt or solvate or stereoisomerthereof.

In yet another of its composition aspects, this invention is directed toa compound of formula III:

wherein

R⁴ is as defined herein (including any specific or preferredembodiments);

W represents O or NH;

or a pharmaceutically acceptable salt or solvate or stereoisomerthereof.

In still another of its composition aspects, this invention is directedto a compound of formula IV:

wherein

R⁴ is as defined herein (including any specific or preferredembodiments);

W represents O or NH;

or a pharmaceutically acceptable salt or solvate or stereoisomerthereof.

In another of its composition aspects, this invention is directed to apharmaceutical composition comprising a pharmaceutically acceptablecarrier and a therapeutically effective amount of a compound of formulaI or a pharmaceutically acceptable salt or solvate or stereoisomerthereof. Such pharmaceutical compositions may optionally contain othertherapeutic agents. Accordingly, in one embodiment, this invention isdirected to such a pharmaceutical composition wherein the compositionfurther comprises a therapeutically effective amount of a steroidalanti-inflammatory agent, such as a corticosteroid.

Compounds of this invention possess both β₂ adrenergic receptor agonistactivity and muscarinic receptor antagonist activity. Accordingly, thecompounds of formula I are useful for treating pulmonary disorders, suchas asthma and chronic obstructive pulmonary disease.

Accordingly, in one of its method aspects, this invention is directed toa method for treating a pulmonary disorder, the method comprisingadministering to a patient in need of treatment a therapeuticallyeffective amount of a compound of formula I or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof.

Additionally, in another of its method aspects, this invention isdirected to a method of providing bronchodilation in a patient, themethod comprising administering to a patient requiring bronchodilation atherapeutically effective amount of a compound of formula I or apharmaceutically acceptable salt or solvate or stereoisomer thereof.

This invention is also directed to a method of treating chronicobstructive pulmonary disease or asthma, the method comprisingadministering to a patient in need of treatment a therapeuticallyeffective amount of a compound of formula I or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof.

Since compounds of this invention possess both β₂ adrenergic receptoragonist activity and muscarinic receptor antagonist activity, suchcompounds are also useful as research tools. Accordingly, in yet anotherof its method aspects, this invention is directed to a method for usinga compound of formula I or a pharmaceutically acceptable salt or solvateor stereoisomer thereof as a research tool for studying a biologicalsystem or sample, or for discovering new chemical compounds having bothβ₂ adrenergic agonist activity and muscarinic receptor antagonistactivity.

This invention is also directed to processes and novel intermediatesuseful for preparing compounds of formula I or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof. Accordingly, inanother of its method aspects, this invention is directed to a processof preparing a compound of formula I, the process comprising:

(a) reacting a compound of formula 1 or a salt thereof, with a compoundof formula 2;

(b) reacting a compound of formula 3 or a salt thereof, with a compoundof formula 4;

(c) coupling a compound of formula 5 with a compound of formula 6;

(d) for a compound of formula I wherein R⁵ represents a hydrogen atom,reacting a compound of formula 3 with a compound of formula 7 or ahydrate thereof, in the presence of a reducing agent;

(e) reacting a compound of formula 1 with a compound of formula 8 or ahydrate thereof, in the presence of a reducing agent;

(f) reacting a compound of formula 9, with a compound of formula 10; or

(g) reacting a compound of formula 11 or a hydrate thereof, with acompound of formula 10, in the presence of a reducing agent;

and then removing any protecting groups to form a compound of formula I;wherein the compounds of formula 1-11 are as defined therein.

In one embodiment, the above process further comprises the step offorming a pharmaceutically acceptable salt of a compound of formula I.In other embodiments, this invention is directed to the other processesdescribed herein; and to the product prepared by any of the processesdescribed herein.

This invention is also directed to a compound of formula I or apharmaceutically acceptable salt or solvate or stereoisomer thereof, foruse in therapy or as a medicament.

Additionally, this invention is directed to the use of a compound offormula I or a pharmaceutically acceptable salt or solvate orstereoisomer thereof, for the manufacture of a medicament; especiallyfor the manufacture of a medicament for the treatment of a pulmonarydisorder.

DETAILED DESCRIPTION OF THE INVENTION

In one of its composition aspects, this invention is directed to novelbiphenyl derivatives of formula I or pharmaceutically acceptable saltsor solvates or stereoisomers thereof. These compounds contain one ormore chiral centers and therefore, this invention is directed to racemicmixtures; pure stereoisomers (i.e., enantiomers or diastereomers);stereoisomer-enriched mixtures and the like unless otherwise indicated.When a particular stereoisomer is shown or named herein, it will beunderstood by those skilled in the art that minor amounts of otherstereoisomers may be present in the compositions of this inventionunless otherwise indicated, provided that the utility of the compositionas a whole is not eliminated by the presence of such other isomers.

In particular, compounds of formula I contain a chiral center at thecarbon atom indicated by the symbol * in the following formula:

In one embodiment of this invention, the carbon atom identified by thesymbol * has the (R) configuration. In this embodiment, it is preferredfor compounds of formula I to have the (R) configuration at the carbonatom identified by the symbol * or to be enriched in a stereoisomericform having the (R) configuration at this carbon atom. In anotherembodiment of this invention, the carbon atom identified by the symbol *has the (S) configuration. In this embodiment, it is preferred forcompounds of formula I to have the (S) configuration at the carbon atomidentified by the symbol * or to be enriched in a stereoisomeric formhaving the (S) configuration at this carbon atom. In some cases, inorder to optimize the β₂ adrenergic agonist activity of the compounds ofthis invention, it is preferred that the carbon atom identified by thesymbol * has the (R) configuration.

The compounds of formula I also contain several basic groups (e.g.,amino groups) and therefore, the compounds of formula I can exist as thefree base or in various salt forms. All such salt forms are includedwithin the scope of this invention. Furthermore, solvates of compoundsof formula I or salts thereof are included within the scope of thisinvention.

Additionally, where applicable, all cis-trans or E/Z isomers (geometricisomers), tautomeric forms and topoisomeric forms of the compounds offormula I are included within the scope of this invention unlessotherwise specified.

The nomenclature used herein to name the compounds of this invention andintermediates thereof has generally been derived using thecommercially-available AutoNom software (MDL, San Leandro, Calif.).Typically, compounds of formula I wherein W is O have been named asester derivatives of biphenyl-2-ylcarbamic acid; and compounds offormula I wherein W is NW^(a) have been named as urea derivatives.

REPRESENTATIVE EMBODIMENTS

The following substituents and values are intended to providerepresentative examples of various aspects and embodiments of thisinvention. These representative values are intended to further defineand illustrate such aspects and embodiments and are not intended toexclude other embodiments or to limit the scope of this invention. Inthis regard, the representation that a particular value or substituentis preferred is not intended in any way to exclude other values orsubstituents from this invention unless specifically indicated.

In particular embodiments of the compounds of formula 1, a and b areindependently 0, 1 or 2; including 0 or 1. In one embodiment, both a andb are 0.

When present, each R¹ may be at the 2, 3, 4, 5 or 6-position of thephenyl ring to which it is attached. In one embodiment, each R¹ isindependently selected from (1-4C)alkyl, halo, —OR^(1a) and—NR^(1f)R^(1g); such as methyl, fluoro, chloro, bromo, hydroxy, methoxy,amino, methylamino, dimethylamino and the like. Particular values for R¹are fluoro or chloro.

When present, each R² may be at the 3, 4, 5 or 6-position on thephenylene ring to which it is attached (where the carbon atom on thephenylene ring attached to the nitrogen atom is position 1). In oneembodiment, each R² is independently selected from (1-4C)alkyl, halo,—OR^(2a) and —NR^(2f)R^(2g); such as methyl, fluoro, chloro, bromo,hydroxy, methoxy, amino, methylamino, dimethylamino and the like.Particular values for R² are fluoro or chloro.

Each R^(1a), R^(1b), R^(1c), R^(1d), R^(1e), R^(1f) and R^(1g) andR^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g) as used in R¹and R², respectively, is independently hydrogen, (1-4C)alkyl orphenyl-(1-4C)alkyl; such as hydrogen, methyl, ethyl, n-propyl,isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl or benzyl. In oneembodiment, these groups are independently hydrogen or (1-3C)alkyl. Inanother embodiment, these groups are independently hydrogen, methyl orethyl.

In one embodiment of this invention, W is O. In another embodiment, W isNW^(a).

Generally, it has been found that compounds in which W represents Oexhibit particularly high affinity for muscarinic and β₂ adrenergicreceptors. Accordingly, in a particular embodiment of this invention, Wpreferably represents O.

When referring to W, particular mention may be made of compounds whereinW is attached to the piperidine ring at the 4-position with respect tothe nitrogen atom of the piperidine ring.

When W is NW^(a), W^(a) is hydrogen or (1-4C)alkyl; such as hydrogen,methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl andtert-butyl. In one embodiment, W^(a) is hydrogen or (1-3C)alkyl. Inanother embodiment, W^(a) is hydrogen, methyl or ethyl; such as hydrogenor methyl. In yet another embodiment, W^(a) is hydrogen and NW^(a) isNH.

In a particular embodiment of the compounds of formula I, c is 0, 1 or2; including 0 or 1. In one embodiment, c is 0.

In one embodiment, each R³ is at the 3, 4 or 5-position on thepiperidine ring (where the nitrogen atom of the piperidine ring isposition 1). In another embodiment, R³ is at 4-position on thepiperidine ring. In a particular aspect of these embodiments, each R³ isindependently selected from (1-4C)alkyl; such as methyl, ethyl,n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl. Inanother aspect, each R³ is independently methyl or ethyl.

In another embodiment, R³ is at the 1-position of the piperidine ring,i.e., on the nitrogen atom of the piperidine ring thus forming aquaternary amine salt. In a particular aspect of this embodiment, eachR³ is independently selected from (1-4C)alkyl; such as methyl, ethyl,n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl. Inanother aspect, each R³ is independently methyl or ethyl.

In yet another embodiment, two R³ groups are joined to form a(1-3C)alkylene or (2-3C)alkenylene group. For example, two R³ groups atthe 2 and 6-positions on the piperidine ring can be joined to form anethylene bridge (i.e., the piperidine ring and the R³ groups form an8-azabicyclo[3.2.1]octane ring); or two R³ groups at the 1 and4-positions on the piperidine ring can be joined to form an ethylenebridge (i.e., the piperidine ring and the R³ groups form an1-azabicyclo[2.2.2]octane ring). In this embodiment, other R³ groups asdefined herein may also be present.

In still another embodiment, two R³ groups are joined to form aoxiran-2,3-diyl group. For example, two R³ groups at the 2 and6-positions on the piperidine ring can be joined to form a3-oxatricyclo[3.3.1.0^(2,4)]nonane ring). In this embodiment, other R³groups as defined herein may also be present.

Each R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3f) and R^(3g) as usedin R³ is independently hydrogen or (1-4C)alkyl; such as hydrogen,methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl andtert-butyl. In one embodiment, these groups are independently hydrogenor (1-3C)alkyl. In another embodiment, these groups are independentlyhydrogen, methyl or ethyl.

In one embodiment of the compounds of formula I, R⁵ is hydrogen or(1-4C)alkyl; such as hydrogen, methyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl and tert-butyl. In another embodiment, eachR⁵ is independently hydrogen, methyl or ethyl. In a particularembodiment, R⁵ is hydrogen.

In one embodiment of this invention, R⁶ is —NR^(6a)CR^(6b)(O) and R⁷ ishydrogen, where each of R^(6a) and R^(6b) is independently hydrogen or(1-4C)alkyl, such as hydrogen, methyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl and tert-butyl. In one embodiment, thesegroups are independently hydrogen or (1-3C)alkyl. In another embodiment,these groups are independently hydrogen, methyl or ethyl. A particularvalue for R⁶ in this embodiment is —NHCHO.

In another embodiment, R⁶ and R⁷ together form—NR^(7a)C(O)—CR^(7b)═CR^(7c)—, —CR^(7d)═CR^(7e)—C(O)—NR^(7f)—,—NR^(7g)C(O)—CR^(7h)R^(7i)—CR^(7j)R^(7k) or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p); where each of R^(7a), R^(7b),R^(7c), R^(7d), R^(7e), R^(7f), R^(7g), R^(7h), R^(7i), R^(7j), R^(7k),R^(7l), R^(7m), R^(7n), R^(7o) and R^(7p) is independently hydrogen or(1-4C)alkyl; such as hydrogen, methyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl and tert-butyl. In one embodiment, thesegroups are independently hydrogen or (1-3C)alkyl. In another embodiment,these groups are independently hydrogen, methyl or ethyl. Particularvalues for R⁶ and R⁷ in this embodiment are R⁶ and R⁷ together form—NHC(O)—CH═CH—, —CH═CH—C(O)—NH—, —CH₂—CH₂—C(O)NH— or —NHC(O)—CH₂—CH₂—;including where R⁶ and R⁷ together form —NHC(O)—CH═CH— or—CH═CH—C(O)—NH—; and in particular, where R⁶ and R⁷ together form—NHC(O)—CH═CH— (i.e., the nitrogen atom is attached at R⁶ and the carbonatom is attached at R⁷ to form, together with the hydroxyphenyl ring towhich R⁶ and R⁷ are attached, a 8-hydroxy-2-oxo-1,2-dihydroquinolin-5-ylgroup).

In the compounds of formula I, R⁴ is a divalent group of the formula:—(R^(4a))_(d)-(A¹)_(e)-(R^(4b))_(f)-Q-(R^(4c))_(g)-(A²)_(h)-(R^(4d))_(i)—

wherein R⁴, A¹, R^(4b), Q, R^(4c), A², R^(4d), d, e, f, g h and i are asdefined herein. In the compound of this invention, the values of each ofthe components R^(4a), A¹, R^(4b), Q, R^(4c), A² and R^(4d) are selectedsuch that the number of contiguous atoms in the shortest chain betweenthe two nitrogen atoms to which R⁴ is attached is in the range of from 4to 16, (specifically, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16);including 8, 9, 10, 11, 12, 13 or 14; such as 8, 9, 10 or 11; or 9 or10. When selecting values for each variable in R⁴, it will beappreciated by those skilled in the art that values should be selectedsuch that a chemically stable group is formed.

When determining the number of contiguous atoms in the shortest chainbetween the two nitrogen atoms to which R⁴ is attached, each contiguousatom of the chain is counted consecutively starting from the first atomin the R⁴ group adjacent to the nitrogen of the piperidine ring endingwith the last atom in the R⁴ group adjacent to the nitrogen of theaminohydroxyethyl group. Where two or more chains are possible, theshortest chain is used to determine the number of contiguous atoms. Asshown below, for example, when R⁴ is —(CH₂)₂—NHC(O)—CH₂—(phen-1,4-ylene)-CH₂—, there are 10 contiguous atoms in the shortestchain counted consecutively starting from the first atom in the R⁴ groupadjacent to the nitrogen of the piperidine ring ending with the lastatom in the R⁴ group adjacent to the nitrogen of the aminohydroxyethylgroup as shown below:

In one embodiment of R⁴, R^(4a) is selected from (1-10C)alkylene,(2-10C)alkenylene and (2-10C)alkynylene wherein the alkylene group isunsubstituted or substituted with 1 or 2 substituents independentlyselected from (1-4C)alkyl, hydroxy and phenyl. Representative examplesof particular values for R^(4a) are —(CH₂)₂—, —(CH₂)₃—, —(CH₂)₄—,—(CH₂)₅—, —(CH₂)₆—, —(CH₂)₇—, —(CH₂)₈—, —(CH₂)₉—, —(CH₂)₁₀—,—(CH₂)CH(CH₃)—, —(CH₂)C(CH₃)₂—, and —(CH₂)₂C(phenyl)₂-. In anotheraspect, R^(4a) is —(CH₂)C(═CH₂)—.

In one embodiment, d is 1.

In one embodiment, A¹ is an optionally substituted (3-7C)cycloalkylenegroup; including a cyclohexylene group, such as cyclohex-1,4-ylene andcyclohex-1,3-ylene; and a cyclopentylene group, such ascyclopent-1,3-ylene.

In another embodiment, A¹ is an optionally substituted (6-10C)arylenegroup, including a phenylene group, such as phen-1,4-ylene,phen-1,3-ylene and phen-1,2-ylene; and a naphthylene group, such asnaphth-1,4-ylene and napth-1,5-ylene.

In yet another embodiment, A¹ is an optionally substituted(2-9C)heteroarylene group, including a pyridylene group, such aspyrid-1,4-ylene; a furylene group, such as fur-2,5-ylene andfur-2,4-ylene; a thienylene group, such as thien-2,5-ylene andthien-2,4-ylene; and a pyrrolylene, such as pyrrol-2,5-ylene andpyrrol-2,4-ylene.

In still another embodiment, A¹ is an optionally substituted(3-6C)heterocyclene group, including a piperidinylene group, such aspiperidin-1,4-ylene; and a pyrrolidinylene group, such aspyrrolidin-2,5-ylene.

In a particular embodiment, A¹ is an optionally substituted phenylene,thienylene, cyclopentylene, cyclohexylene or piperidinylene.

In one embodiment, e is 0.

In a particular embodiment, R^(4b) is (1-5C)alkylene. Representativeexamples of particular values for R^(4b) are —CH₂—, —(CH₂)₂—, —(CH₂)₃—,—(CH₂)₄—, —(CH₂)₅—; including methylene, ethylene and propylene.

In one embodiment, f is 0.

In a particular embodiment, Q is selected from a bond, —N(Q^(a))C(O)—,—C(O)N(Q^(b))-, —N(Q^(c))S(O)₂—, —S(O)₂N(Q^(d))-,—N(Q^(f))C(O)N(Q^(f))-, —OC(O)N(Q^(i))-, —N(Q^(j))C(O)O— or —N(Q^(k));such as where Q is a bond, —N(Q^(a))C(O)— or —C(O)N(Q^(b))—.Representative examples of particular values for Q are a bond, O, NH,—C(O)NH—, —C(O)N(CH₃)—, —NHC(O)—, —N(CH₃)C(O)—, —S(O)₂NH—,—S(O)₂N(CH₃)—, —NHS(O)₂—, —N(CH₃)S(O)₂— and —NHC(O)NH—. Another exampleof a value for Q, together with R^(4c), is —C(O)(piperidin-1,4-ylene).

In one embodiment, Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g),Q^(h), Q^(j), Q^(i) and Q^(k) are each independently selected fromhydrogen and (1-6C)alkyl, wherein the alkyl group is unsubstituted orsubstituted with from 1 to 3 substituents independently selected fromfluoro, hydroxy and (1-4C)alkoxy. For example, Q^(a), Q^(b), Q^(c),Q^(d), Q^(e), Q^(f), Q^(g), Q^(h), Q^(i), Q^(j) and Q^(k) are eachindependently selected from hydrogen, and (1-3C)alkyl, includinghydrogen, methyl, ethyl, n-propyl and isopropyl. An example of a valuefor each of Q^(a), Q^(b), Q^(c), Q^(d), Q^(c), Q^(f), Q^(g), Q^(h),Q^(i), Q^(j) and Q^(k) is hydrogen.

In another embodiment, Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g),Q^(h), Q^(i), Q^(j) and Q^(k) together with the nitrogen atom and thegroup R^(4b) or R^(4c) to which they are attached, form a 4-6 memberedazacycloalkylene group. For example, Q^(a) and Q^(b) together with thenitrogen atom and the group R^(4b) or R^(4c) to which they are attached,form a piperidin-4-ylene group. By way of illustration, when Qrepresents —N(Q^(a))C(O)— and Q^(a) together with the nitrogen atom andthe group R^(4b) to which it is attached, forms a piperidin-4-ylenegroup, R⁴ is a group of formula:

Similarly, when Q represents —C(O)N(Q^(b))- and Q^(b) together with thenitrogen atom and the group R^(4c) to which it is attached, forms apiperidin-4-ylene group, R⁴ is a group of formula:

In a particular embodiment, R^(4c) is (1-5C)alkylene. Representativeexamples of particular values for R^(4c) are —CH₂—, —(CH₂)₂—, —(CH₂)₃—,—(CH₂)₄—, —(CH₂)₅—; including methylene, ethylene and propylene.

In one embodiment, A² is an optionally substituted (3-7C)cycloalkylenegroup; including a cyclohexylene group, such as cyclohex-1,4-ylene andcyclohex-1,3-ylene; and a cyclopentylene group, such ascyclopent-1,3-ylene.

In another embodiment, A² is an optionally substituted (6-10C)arylenegroup, including a phenylene group, such as phen-1,4-ylene,phen-1,3-ylene and phen-1,2-ylene; and a naphthylene group, such asnaphth-1,4-ylene and napth-1,5-ylene.

In yet another embodiment, A² is an optionally substituted(2-9C)heteroarylene group, including a pyridylene group, such aspyrid-1,4-ylene; a furylene group, such as fur-2,5-ylene andfur-2,4-ylene; a thienylene group, such as thien-2,5-ylene andthien-2,4-ylene; and a pyrrolylene, such as pyrrol-2,5-ylene andpyrrol-2,4-ylene.

In still another embodiment, A² is an optionally substituted(3-6C)heterocyclene group, including a piperidinylene group, such aspiperidin-1,4-ylene; and a pyrrolidinylene group, such aspyrrolidin-2,5-ylene.

In a particular embodiment, A² is optionally substituted phenylene,thienylene, cyclopentylene, cyclohexylene or piperidinylene.

By way of illustration, either A¹ or A² or both can be phenylene, suchas phen-1,4-ylene or phen-1,3-ylene, where the phenylene group isunsubstituted or substituted with from 1 to 4 substituents independentlyselected from halo, (1-4C)alkyl, (1-4C)alkoxy, —S-(1-4C)alkyl,—S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl, —C(O)O(1-4C)alkyl, carboxy,cyano, hydroxy, nitro, trifluoromethyl and trifluoromethoxy.Representative examples include phen-1,3-ylene, phen-1,4-ylene,4-chlorophen-1,3-ylene, 6-chlorophen-1,3-ylene, 4-methylphen-1,3-ylene,2-fluorophen-1,4-ylene, 2-chlorophen-1,4-ylene, 2-bromophen-1,4-ylene,2-iodophen-1,4-ylene, 2-methylphen-1,4-ylene, 2-methoxyphen-1,4-ylene,2-trifluoromethoxyphen-1,4-ylene, 3-nitrophen-1,4-ylene,3-chlorophen-1,4-ylene, 2,5-difluorophen-1,4-ylene,2,6-dichlorophen-1,4-ylene, 2,6-diiodophen-1,4-ylene,2-chloro-6-methylphen-1,4-ylene, 2-chloro-5-methoxyphen-1,4-ylene,2,3,5,6-tetrafluorophen-1,4-ylene.

Alternatively, A¹ or A² or both can be cyclopentylene or cyclohexylene;wherein the cyclopentylene or cyclohexylene group is unsubstituted orsubstituted with (1-4C)alkyl. Representative examples includecis-cyclopent-1,3-ylene, trans-cyclopent-1,3-ylene,cis-cyclohex-1,4-ylene and trans-cyclohex-1,4-ylene. A¹ or A² or bothcan also be optionally substituted thienylene or piperidinylene, forexample, thien-2,5-ylene or piperidin-1,4-ylene.

In one embodiment, R^(4d) is selected from (1-10C)alkylene,(2-10C)alkenylene and (2-10C)alkynylene wherein the alkylene isunsubstituted or substituted with 1 or 2 substituents independentlyselected from (1-4C)alkyl, hydroxy and phenyl. Representative examplesof particular values for R^(4d) are —(CH₂)—, —(CH₂)₂—, —(CH₂)₃—,—(CH₂)₄—, —(CH₂)₅—, —(CH₂)₆—, —(CH₂)₇—, —(CH₂)₈—, —(CH₂)₉—, —(CH₂)₁₀—and —(CH₂)CH(CH₃)—(CH₂)—C(CH₃)₂—(CH₂)₂—.

In a particular embodiment, R⁴ is a divalent group of the formula:—(R^(4a))_(d)— where R^(4a) is (4-10C)alkylene. In one aspect of thisembodiment, R⁴ is a divalent group of the formula: —(CH₂)_(j)— where jis 8, 9 or 10. Examples of particular values for R⁴ in this embodimentare —(CH₂)₄—, —(CH₂)₅—, —(CH₂)₆—, —(CH₂)₇—, —(CH₂)₈—, —(CH₂)₉, and—(CH₂)₁₀—; including —(CH₂)₈—, —(CH₂)₉, and —(CH₂)₁₀—.

In another particular embodiment, R⁴ is a divalent group of the formula:—(R^(4a))_(d)-(A²)_(h)-(R^(4d))_(i)—

where R^(4a) is (1-10C)alkylene, such as —(CH₂)—, —(CH₂)₂—, —(CH₂)₃—; A²is (6-10C)arylene, such as phen-1,4-ylene or phen-1,3-ylene, or(2-9C)heteroarylene, such as thien-2,5-ylene or thien-2,4-ylene; andR^(4d) is (1-10C)alkylene, such as —(CH₂)—, —(CH₂)₂—, —(CH₂)₃—. Examplesof particular values for R⁴ in this embodiment are—(CH₂)-(phen-1,4-ylene)-(CH₂)—; —(CH₂)-(phen-1,4-ylene)-(CH₂)₂—;—(CH₂)-(phen-1,4-ylene)-(CH₂)₃—; —(CH₂)₂-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₂-(phen-1,4-ylene)-(CH₂)₂—; —(CH₂)₂-(phen-1,4-ylene)-(CH₂)₃—;—(CH₂)₃-(phen-1,4-ylene)-(CH₂)—; —(CH₂)₃-(phen-1,4-ylene)-(CH₂)₂—,—(CH₂)₃-(phen-1,4-ylene)-(CH₂)₃—, —(CH₂)₄-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₄-(phen-1,4-ylene)-(CH₂)₂— and —(CH₂)₄-(phen-1,4-ylene)-(CH₂)₃—.

In yet another particular embodiment, R⁴ is a divalent group of theformula:—(R^(4a))_(d)-Q-(A²)_(h)-(R^(4d))_(i)—

where Q is —O— or —N(Q^(k))-; Q^(k) is hydrogen or (1-3C)alkyl, such asmethyl or ethyl; R^(4a) is (1-10C)alkylene, such as —(CH₂)—, —(CH₂)₂—,—(CH₂)₃—; A² is (6-10C)arylene, such as phen-1,4-ylene orphen-1,3-ylene, or (2-9C)heteroarylene, such as thien-2,5-ylene orthien-2,4-ylene; and R^(4d) is (1-10C)alkylene, such as —(CH₂)—,—(CH₂)₂—, —(CH₂)₃—. Examples of particular values for R⁴ in thisembodiment are —(CH₂)₂—O-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₂—O-(phen-1,4-ylene)-(CH₂)₂—; —(CH₂)₂—O-(phen-1,4-ylene)-(CH₂)₃—;—(CH₂)₃—O-(phen-1,4-ylene)-(CH₂)—; —(CH₂)₃—O-(phen-1,4-ylene)-(CH₂)₂—;—(CH₂)₃—O-(phen-1,4-ylene)-(CH₂)₃—; —(CH₂)₂—NH-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₂—NH-(phen-1,4-ylene)-(CH₂)₂—;—(CH₂)₂—NH-(phen-1,4-ylene)-(CH₂)₃—; —(CH₂)₃—NH-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₃—NH-(phen-1,4-ylene)-(CH₂)₂— and—(CH₂)₃—NH-(phen-1,4-ylene)-(CH₂)₃—.

In yet another particular embodiment, R⁴ is a divalent group of theformula:—(R^(4a))_(d)-(A¹)_(e)-(R^(4b))_(f)-Q-(R^(4c))_(g)-(A²)_(h)-(R^(4d))_(i)—

where Q is —N(Q^(a))C(O)— or —C(O)N(Q^(b))-. A particular value for R⁴in this embodiment is the formula:

where m is an integer from 2 to 10; and n is an integer from 2 to 10;provided that m+n is an integer from 4 to 12. In this formula for R⁴, dand g are 1 and e, f, h and i are 0; and R^(4a) is —(CH₂)_(m)—, R^(4c)is —(CH₂)_(n)— and Q is —C(O)NH—. Particular values for m are 2 or 3;and for n, 4, 5 or 6.

Another particular value for R⁴ is the formula:

where o is an integer from 2 to 7; and p is an integer from 1 to 6;provided that o+p is an integer from 3 to 8. In this formula for R⁴, d,h and i are 1 and e, f and g are 0; and R^(4a) is —(CH₂)_(o)—, A² isphen-1,4-ylene, R^(4d) is —(CH₂)_(p)— and Q is —C(O)NH—. Particularvalues for o are 2 or 3; and for p, 1 or 2. In this embodiment, thephen-1,4-ylene group may be optionally substituted as defined herein forA².

Another particular value for R⁴ is the formula:

where q is an integer from 2 to 6; r is an integer from 1 to 5; and s isan integer from 1 to 5; provided that q+r+s is an integer from 4 to 8.In this formula for R⁴, d, g, h and i are 1 and e and f are 0; andR^(4a) is —(CH₂)_(q)—, R^(4c) is —(CH₂)_(r)—, A² is 1,4-phenylene,R^(4d) is —(CH₂)_(s)— and Q is —C(O)NH—. Particular values for q are 2or 3; for r, 1 or 2; and for s, 1 or 2. In this embodiment, thephen-1,4-ylene group may be optionally substituted as defined herein forA².

Another particular value for R⁴ is the formula:

where t is an integer from 2 to 10; and u is an integer from 2 to 10;provided that t+u is an integer from 4 to 12. In this formula for R⁴, dand g are 1 and e, f, h and i are 0; and R^(4a) is —(CH₂)_(t)—, R^(4c)is —(CH₂)_(u)— and Q is —NHC(O)—. Particular values for t are 2 or 3;and for u, 4, 5 or 6.

Another particular value for R⁴ is the formula:

where v is an integer from 2 to 7; and w is an integer from 1 to 6;provided that v+w is an integer from 3 to 8. In this formula for R⁴, d,h and i are 1 and e, f and g are 0; and R^(4a) is —(CH₂)_(v)—, A² is1,4-phenylene, R^(4d) is —(CH₂)_(w)— and Q is —NHC(O)—. Particularvalues for v are 2 or 3; and for w, 1 or 2. In this embodiment, thephen-1,4-ylene group may be optionally substituted as defined herein forA².

Another particular value for R⁴ is the formula:

where x is an integer from 2 to 6; y is an integer from 1 to 5; and z isan integer from 1 to 5; provided that x+y+z is an integer from 4 to 8.In this formula for R⁴, d, g, h and i are 1 and e and f are 0; andR^(4a) is —(CH₂)_(x)—, R^(4c) is —(CH₂)_(y)—, A² is 1,4-phenylene,R^(4d) is —(CH₂)_(z)— and Q is —NHC(O)—. Particular values for x are 2or 3; for y, 1 or 2; and for z, 1 or 2. In this embodiment, thephen-1,4-ylene group may be optionally substituted as defined herein forA².

By way of further illustration, R⁴ can be selected from:

—(CH₂)₇—;

—(CH₂)₈—;

—(CH₂)₉—;

—(CH₂)₁₀—;

—(CH₂)₁₁—;

—(CH₂)₂C(O)NH(CH₂)₅—;

—(CH₂)₂N(CH₃)C(O)(CH₂)₅—;

—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂—;

—(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂—;

—(CH₂)₂NHC(O)NH(CH₂)₅—;

—(CH₂)₃NHC(O)NH(CH₂)₅—;

—(CH₂)₂C(O)NHCH₂(cyclohex-1,3-ylene)CH₂—;

—(CH₂)₂NHC(O)(cyclopent-1,3-ylene)-;

—(CH₂)₂NHC(O)NH(phen-1,4-ylene)(CH₂)₂—;

1-[—(CH₂)₂C(O)](piperidin-4-yl)(CH₂)₂—;

—(CH₂)₂NHC(O)(trans-cyclohex-1,4-ylene)CH₂—;

—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)-;

—(CH₂)₂NH(phen-1,4-ylene)(CH₂)₂—;

1-[—(CH₂)₂NHC(O)](piperidin-4-yl)(CH₂)₂—;

—CH₂(phen-1,4-ylene)NH(phen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)CH₂—;

—(CH₂)₂C(O)NHCH₂(pyrid-2,6-ylene)CH₂—;

—(CH₂)₂C(O)NH(cis-cyclohex-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(trans-cyclohex-1,4-ylene)CH₂—;

—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)CH₂—;

—(CH₂)₂N(CH₃)C(O)(phen-1,3-ylene)CH₂—;

—(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)—((S)-isomer);

—(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)—((R)-isomer);

2-[(S)-(—CH₂—](pyrrolidin-1-yl)C(O)(CH₂)₄—;

2-[(S)-(—CH₂—](pyrrolidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(4-chlorophen-1,3-ylene)CH₂—;

—CH₂(2-fluorophen-1,3-ylene)CH₂—;

—(CH₂)₂C(O)NH(4-methylphen-1,3-ylene)CH₂—;

—(CH₂)₂C(O)NH(6-chlorophen-1,3-ylene)CH₂—;

—(CH₂)₂C(O)NH(2-chlorophen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(2,6-dichlorophen-1,4-ylene)CH₂—;

—(CH₂)₂NHC(O)NHCH₂(phen-1,3-ylene)CH₂—;

4-[—CH₂—](piperidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)N(CH₂CH₃)(phen-1,4-ylene)CH₂—;

1-[—(CH₂)₂NHC(O)](piperidin-4-yl)-;

—(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₂NHC(O)(thien-2,5-ylene)CH₂—;

—(CH₂)₂N(CH₃)C(O)(3-nitrophen-1,4-ylene)CH₂—;

—(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)-;

1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)-;

5-[—(CH₂)₂NHC(O)](pyrid-2-yl)CH₂—;

—(CH₂)₂(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₃(thien-2,5-ylene)(CH₂)₃—;

—(CH₂)₂(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

—CH₂(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)(CH₂)₂—;

1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)CH₂—;

—(CH₂)₂C(O)NH(3-chlorophen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(2-(CF₃O—)phen-1,4-ylene)CH₂—;

—(CH₂)₃(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₂S(O)₂NH(CH₂)₅—;

—CH₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₂C(O)NH(2-iodophen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(2-chloro-5-methoxyphen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(2-chloro-6-methylphen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(CH₂)₅—;

—(CH₂)₂N(CH₃)S(O)₂(phen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(2-bromophen-1,4-ylene)CH₂—;

—(CH₂)₃(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₃(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)(CH₂)₃—;

—(CH₂)₂C(O)NH(2-methoxyphen-1,4-ylene)CH₂—;

—(CH₂)₅NH(phen-1,4-ylene)(CH₂)₂—;

4-[—(CH₂)₂-](piperidin-1-yl)(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₂C(O)NH(phen-1,4-ylene)CH(CH₃)CH₂—;

—(CH₂)₂-(trans-cyclohex-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₂C(O)NH(2-fluorophen-1,4-ylene)CH₂—;

—(CH₂)₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₂C(O)NH(2,5-difluorophen-1,4-ylene)CH₂—;

—(CH₂)₂NHC(O)(phen-1,4-ylene)(CH₂)₂—;

1-[—CH₂(pyrid-2,6-ylene)CH₂](piperidin-4-yl)CH₂—;

—(CH₂)₃NH(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₂NH(naphth-1,4-ylene)(CH₂)₂—;

—(CH₂)₃O(phen-1,4-ylene)CH₂—;

1-[—(CH₂)₃](piperidin-4-yl)CH₂—;

4-[—(CH₂)₂](piperidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;

—(CH₂)₃(phen-1,4-ylene)NHC(O)(CH₂)₂—;

—(CH₂)₃O(phen-1,4-ylene)(CH₂)₂—;

2-[—(CH₂)₂](benzimidazol-5-yl)CH₂—;

—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₂—;

—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₄—;

—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₅—;

4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₂—;

—(CH₂)₂NHC(O)NH(phen-1,4-ylene)CH₂—;

—(CH₂)₂N(CH₃)(CH₂)₂(cis-cyclohex-1,4-ylene)-;

—(CH₂)₂C(O)NH(2,3,5,6-tetrafluorophen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(2,6-diiodophen-1,4-ylene)CH₂—;

4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₃—;

4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₄—;

4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₅—;

—(CH₂)₂C(O)NHCH₂(phen-1,4-ylene)CH₂—;

—(CH₂)₂NHC(O)NHCH₂(phen-1,4-ylene)CH₂—;

—(CH₂)₂C(O)NH(2-methylphen-1,4-ylene)CH₂—;

1-[—(CH₂)₃O(phen-1,4-ylene)(CH₂)₂](piperidin-4-yl)CH₂—;

—(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)(CH₂)₂—;

—(CH₂)₂O(phen-1,3-ylene)CH₂—;

—(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,4-ylene)CH₂—;

—(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,3-ylene)CH₂—;

—(CH₂)₂N(CH₃)C(O)(fur-2,5-ylene)CH₂—;

—(CH₂)₂N(CH₃)C(O)(thien-2,5-ylene)CH₂—;

—(CH₂)₂O(phen-1,4-ylene)O(CH₂)₂—;

—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,4-ylene)CH₂—;

—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,2-ylene)CH₂—;

—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,3-ylene)CH₂—;

—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,4-ylene)CH₂—;

—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(fur-2,5-ylene)CH₂—;

—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(thien-2,5-ylene)CH₂—;

4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,2-ylene)CH₂—;

4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,3-ylene)CH₂—;

4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,4-ylene)CH₂—;

4-[—(CH₂)₂](piperidin-1-yl)C(O)(fur-2,5-ylene)CH₂—;

4-[—(CH₂)₂](piperidin-1-yl)C(O)(thien-2,5-ylene)CH₂—;

—(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,3-ylene)CH₂—;

—(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,4-ylene)CH₂—;

—(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,2-ylene)CH₂—;

—(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,3-ylene)CH₂—;

—(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,4-ylene)CH₂—;

—(CH₂)₂(phen-1,4-ylene)NHC(O)(fur-2,5-ylene)CH₂—;

—(CH₂)₂(phen-1,4-ylene)NHC(O)(thien-2,5-ylene)CH₂—;

—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,3-ylene)CH₂—;

—(CH₂)₃O(phen-1,3-ylene)CH₂—;

—CH₂CH(OH)CH₂NH(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₄NH(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂NHC(O)CH₂—;

—(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂NHC(O)CH₂—;

—(CH₂)₂C(O)NHCH₂(trans-cyclohex-1,4-ylene)CH₂—;

—(CH₂)₂NHC(O)(CH₂)₅—;

—(CH₂)₂O(phen-1,3-ylene)O(CH₂)₂—;

—(CH₂)₂O(phen-1,2-ylene)O(CH₂)₂—;

—CH₂(phen-1,2-ylene)O(phen-1,2-ylene)CH₂—;

—(CH₂)₂C(O)NH(CH₂)₆—;

—(CH₂)₃(phen-1,4-ylene)(CH₂)₃—;

—(CH₂)₃(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₄(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₃(furan-2,5-ylene)(CH₂)₃—;

—(CH₂)₂N(CH₃)C(O)NH(phen-1,4-ylene)(CH₂)₂—;

4-[—(CH₂)₂](piperidin-1-yl)C(O)NH(phen-1,4-ylene)(CH₂)₂—;

—(CH₂)₃(phen-1,3-ylene)(CH₂)₃—;

—(CH₂)₃(tetrahydrofuran-2,5-ylene)(CH₂)₃—; and

—(CH₂)₂O(phen-1,4-ylene)C(O)(CH₂)₂—.

Representative Subgeneric Groupings

The following subgeneric formulae and groupings are intended to providerepresentative examples of various aspects and embodiments of thisinvention and as such, they are not intended to exclude otherembodiments or to limit the scope of this invention unless otherwiseindicated.

A particular group of compounds of formula I are those disclosed in U.S.Provisional Application No. 60/447,843, filed on Feb. 14, 2003. Thisgroup includes compounds of formula I; wherein:

a is 0 or an integer of from 1 to 3;

each R¹ is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(1a), —C(O)OR^(1b),SR^(1c), —S(O)R^(1d), S(O)₂R^(1e) and —NR^(1f)R^(1g);

each of R^(1a), R^(1b), R^(1c), R^(1d), R^(1e), R^(1f) and R^(1g) isindependently hydrogen or (1-4C)alkyl;

b is 0 or an integer of from 1 to 3;

each R² is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(2a), C(O)OR^(2b),SR^(2c), S(O)R^(2d), S(O)₂R^(2e) and —NR^(2f)R^(2g);

each of R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g) isindependently hydrogen or (1-4C)alkyl;

W is attached to the 3- or 4-position with respect to the nitrogen atomin the piperidine ring, and represents O or NW^(a);

W^(a) is hydrogen or (1-4C)alkyl;

c is 0 or an integer of from 1 to 4;

each R³ is a substituent on carbon independently selected from(1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano,halo, —OR^(3a), —C(O)OR^(3b), SR^(3c), —S(O)R^(3d), —S(O)₂R^(3e) and—NR^(3f)R^(3g);

each of R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3f) and R^(3g) isindependently hydrogen or (1-4C)alkyl;

R⁴ is a divalent group of the formula:—(R^(4a))_(d)-(A¹)_(e)-(R^(4b))_(f)-Q-(R^(4c))_(g)-(A²)_(h)-(R^(4d))_(i)—wherein

d, e, f, g, h and i are each independently selected from 0 and 1;

R^(4a), R^(4b), R^(4c) and R^(4d) are each independently selected from(1-10C)alkylene, (2-10C)alkenylene and (2-10C)alkynylene wherein eachalkylene, alkenylene or alkynylene group is unsubstituted or substitutedwith from 1 to 5 substituents independently selected from (1-4C)alkyl,fluoro, hydroxy, phenyl and phenyl(1-4C)-alkyl;

A¹ and A² are each independently selected from (3-7C)cycloalkylene,(6-10C)arylene, (2-9C)heteroarylene and (3-6C)heterocyclene; whereineach cycloalkylene is unsubstituted or substituted with from 1 to 4substitutents selected independently from (1-4C)alkyl and each arylene,heteroarylene or heterocyclene group is unsubstituted or substitutedwith from 1 to 4 substituents independently selected from halo,(1-4C)alkyl and (1-4C)alkoxy;

Q is selected from a bond, —O—, —C(O)O—, —OC(O)—, —S—, —S(O)—, —S(O)₂—,—N(Q^(a))C(O)—, —C(O)N(Q^(b))-, —N(Q^(c))S(O)₂—, —S(O)₂N(Q^(d))-,—N(Q^(e))C(O)N(Q^(f))-, —N(Q^(g))S(O)₂N(Q^(h))-, —OC(O)N(Q^(i))- and—N(Q^(j))C(O)O—;

Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g), Q^(h), Q^(i) and Q^(j)are each independently selected from hydrogen, (1-6C)alkyl, A³ and(1-4C)alkylene-A⁴; wherein the alkyl group is unsubstituted orsubstituted with from 1 to 3 substituents independently selected fromfluoro, hydroxy and (1-4C)alkoxy; or together with the nitrogen atom andthe group R^(4b) or R^(4c) to which they are attached, form a 4-6membered azacycloalkylene group;

A³ and A⁴ are each independently selected from (3-6C)cycloalkyl,(6-10C)aryl, (2-9C)heteroaryl and (3-6C)heterocyclyl; wherein eachcycloalkyl is unsubstituted or substituted with from 1 to 4substitutents selected independently from (1-4C)alkyl and each aryl,heteroaryl or heterocyclyl group is unsubstituted or substituted withfrom 1 to 4 substituents independently selected from halo, (1-4C)alkyland (1-4C)alkoxy;

provided that the number of contiguous atoms in the shortest chainbetween the two nitrogen atoms to which R⁴ is attached is in the rangeof from 8 to 14;

R⁵ represents hydrogen or (1-4C)alkyl;

R⁶ is —NR^(6a)CR^(6b)(O) and R⁷ is hydrogen, or R⁶ and R⁷ together form—NR^(7a)C(O)—CR^(7b)═CR^(7c), —CR^(7d)═CR^(7e)—C(O)—NR^(7f)—,—NR^(7g)C(O)—CR^(7h)R⁷—CR^(7j)R^(7k) or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p)—;

each of R^(6a) and R^(6b) is independently hydrogen or (1-4C)alkyl; and

each of R^(7a), R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(7g), R^(7h),R^(7i), R^(7j), R^(7k), R^(7l), R^(7m), R^(7n), R^(7o) and R^(7p) isindependently hydrogen or (1-4C)alkyl; or a pharmaceutically acceptablesalt or solvate or stereoisomer thereof.

Another particular group of compounds of formula I are those disclosedin U.S. Provisional Application No. 60/467,035, filed on May 1, 2003.This group of compounds includes compounds of formula I; wherein:

a is 0 or an integer of from 1 to 3;

each R¹ is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(1a), —C(O)OR^(1b),SR^(1c), —S(O)R^(1d), —S(O)₂R^(1e), and —NR^(1f)R^(1g);

each of R^(1a), R^(1b), R^(1c), R^(1d), R^(1e), R^(1f) and R^(1g) isindependently hydrogen or (1-4C)alkyl;

b is 0 or an integer of from 1 to 3;

each R² is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(2a), —C(O)OR^(2b),SR^(2c), —S(O)R^(2d), —S(O)₂R^(2e), and —NR^(2f)R^(2g);

each of R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g) isindependently hydrogen or (1-4C)alkyl;

W is attached to the 3- or 4-position with respect to the nitrogen atomin the piperidine ring, and represents O or NW^(a);

W^(a) is hydrogen or (1-4C)alkyl;

c is 0 or an integer of from 1 to 4;

each R³ is a substituent on carbon independently selected from(1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano,halo, —OR^(3a), —C(O)OR^(3b), SR^(3c), —S(O)R^(3d), —S(O)₂R^(3e), and—NR^(3f)R^(3g);

each of R^(3a), R^(3b), R^(3c), R^(3d), R^(3e), R^(3f) and R^(3g) isindependently hydrogen or (1-4C)alkyl;

R⁴ is a divalent group of the formula:—(R^(4a))_(d)-(A¹)_(e)-(R^(4b))_(f)-Q-(R^(4c))_(g)-(A²)_(h)-(R^(4d))_(i)—wherein

d, e, f, g, h and i are each independently selected from 0 and 1;

R^(4a), R^(4b), R^(4c) and R^(4d) are each independently selected from(1-10C)alkylene, (2-10C)alkenylene and (2-10C)alkynylene wherein eachalkylene, alkenylene or alkynylene group is unsubstituted or substitutedwith from 1 to 5 substituents independently selected from (1-4C)alkyl,fluoro, hydroxy, phenyl and phenyl(1-4C)-alkyl;

A¹ and A² are each independently selected from (3-7C)cycloalkylene,(6-10C)arylene, (2-9C)heteroarylene and (3-6C)heterocyclene; whereineach cycloalkylene is unsubstituted or substituted with from 1 to 4substitutents selected independently from (1-4C)alkyl and each arylene,heteroarylene or heterocyclene group is unsubstituted or substitutedwith from 1 to 4 substituents independently selected from halo,(1-4C)alkyl and (1-4C)alkoxy;

Q is selected from a bond, —O—, —C(O)O—, —OC(O)—, —S—, —S(O)—, —S(O)₂—,—N(Q^(a))C(O)—, —C(O)N(Q^(b))-, —N(Q^(c))S(O)₂—, —S(O)₂N(Q^(d))-,—N(Q^(e))C(O)N(Q^(f))-, —N(Q^(g))S(O)₂N(Q^(h))-, —OC(O)N(Q^(i))- and—N(Q^(j))C(O)O—;

Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g), Q^(h), Q^(i) and Q^(j)are each independently selected from hydrogen, (1-6C)alkyl, A³ and(1-4C)alkylene-A⁴; wherein the alkyl group is unsubstituted orsubstituted with from 1 to 3 substituents independently selected fromfluoro, hydroxy and (1-4C)alkoxy; or together with the nitrogen atom andthe group R^(4b) or R^(4c) to which they are attached, form a 4-6membered azacycloalkylene group;

A³ and A⁴ are each independently selected from (3-6C)cycloalkyl,(6-10C)aryl, (2-9C)heteroaryl and (3-6C)heterocyclyl; wherein eachcycloalkyl is unsubstituted or substituted with from 1 to 4substitutents selected independently from (1-4C)alkyl and each aryl,heteroaryl or heterocyclyl group is unsubstituted or substituted withfrom 1 to 4 substituents independently selected from halo, (1-4C)alkyland (1-4C)alkoxy;

provided that the number of contiguous atoms in the shortest chainbetween the two nitrogen atoms to which R⁴ is attached is in the rangeof from 4 to 14;

R⁵ represents hydrogen or (1-4C)alkyl;

R⁶ is —NR^(6a)CR^(6b)(O) or CR^(6c)R^(6d)OR^(6e) and R⁷ is hydrogen, orR⁶ and R⁷ together form —NR^(7a)C(O)—CR^(7b)═CR^(7c)—,CR^(7d)═CR^(7e)—C(O)—NR^(7f), —NR^(7g)C(O)CR^(7h)R^(7i)—CR^(7j)R^(7k)—or —CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p)—;

each of R^(6a), R^(6b), R^(6c), R^(6d) and R^(6e) is independentlyhydrogen or (1-4C)alkyl; and

each of R^(7a), R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(7g), R^(7h),R^(7i), R^(7j), R^(7k), R^(7l), R^(7m), R^(7n), R^(7o) and R^(7p) isindependently hydrogen or (1-4C)alkyl;

or a pharmaceutically acceptable salt or solvate or stereoisomerthereof.

Another particular group of compounds of formula I are those where: a is0; b is 0; c is 0; W is O; W is attached at the 4-position of thepiperidinyl ring; R⁵ is hydrogen; and R⁴, R⁶ and R⁷ are as definedherein; or a pharmaceutically acceptable salt or solvate or stereoisomerthereof.

Still another particular group of compounds of formula I are thosewherein: a is 0; b is 0; c is 0; W is NH; W is attached at the4-position of the piperidinyl ring; R⁵ is hydrogen; and R⁴, R⁶ and R⁷are as defined herein; or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

Yet another particular group of compounds of formula I are thosewherein: a is 0; b is 0; c is 0; W is O; W is attached at the 4-positionof the piperidinyl ring; R⁴ is —(CH₂)_(j)— where j is 8, 9 or 10; R⁵ ishydrogen; and R⁶ and R⁷ are as defined herein; or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof.

Another particular group of compounds of formula I are those wherein: ais 0; b is 0; c is 0; W is NH; W is attached at the 4-position of thepiperidinyl ring; R⁴ is —(CH₂)_(j)— where j is 8, 9 or 10; R⁵ ishydrogen; and R⁶ and R⁷ are as defined herein; or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof.

Yet another particular group of compounds of formula I are thosewherein: a is 0; b is 0; c is 0; W is O; W is attached at the 4-positionof the piperidinyl ring; R⁴ is —(CH₂)₂—C(O)NH—(CH₂)₅—; R⁵ is hydrogen;and R⁶ and R⁷ are as defined herein; or a pharmaceutically acceptablesalt or solvate or stereoisomer thereof.

Another particular group of compounds of formula I are those wherein: ais 0; b is 0; c is 0; W is NH; W is attached at the 4-position of thepiperidinyl ring; R⁴ is —(CH₂)₂—C(O)NH—(CH₂)₅—; R⁵ is hydrogen; and R⁶and R⁷ are as defined herein; or a pharmaceutically acceptable salt orsolvate or stereoisomer thereof.

Another particular group of compounds of formula I are those of formulaII as defined herein; or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

Another particular group of compounds of formula I are those of formulaIII as defined herein; or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

Another particular group of compounds of formula I are those of formulaIV as defined herein; or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

Another particular group of compounds of formula I are those of formulaII, III or IV as defined herein, wherein the piperidinyl ring issubstituted at the 4-position with a methyl group; or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof.

Another particular group of compounds of formula I are compounds offormula V:

wherein W, R⁴, R⁶ and R⁷ are as defined in Table I; or apharmaceutically acceptable salt or solvate thereof. TABLE I Ex. W R⁴ R⁶R⁷ 1 NH —(CH₂)₉-(racemic)¹ —NHC(O)CH═CH—² 2 O —(CH₂)₉-(racemic)—NHC(O)CH═CH— 3 O —(CH₂)₉— —NHC(O)CH═CH— 4 O —(CH₂)₉— H —NHC(O)H 5 O—(CH₂)₉— —NHC(O)CH₂CH₂— 6 O —(CH₂)₂C(O)NH(CH₂)₅— —NHC(O)CH═CH— 7 O—(CH₂)₂N(CH₃)C(O)(CH₂)₅— —NHC(O)CH═CH— 8 O—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 9 O—(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 10 O—(CH₂)₂NHC(O)NH(CH₂)₅— —NHC(O)CH═CH— 11 O —(CH₂)₃NHC(O)NH(CH₂)₅——NHC(O)CH═CH— 12 O —(CH₂)₉— H —CH₂OH 13 NH —(CH₂)₉— H —CH₂OH 14 O—(CH₂)₂C(O)NHCH₂(cyclohex-1,3-ylene)CH₂— —NHC(O)CH═CH— 15 O—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)- —NHC(O)CH₂CH₂— 16 O—(CH₂)₂C(O)NH(2-chlorophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 17 O—(CH₂)₂S(O)₂NH(CH₂)₅— —NHC(O)CH═CH— 18 O—(CH₂)₂N(CH₃)S(O)₂(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 19 O—(CH₂)₂NHC(O)NHCH₂(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 20 O—(CH₂)₃(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 21 O1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4- —NHC(O)CH═CH— yl)CH₂—22 O —(CH₂)₃O(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 23 O—(CH₂)₂(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 24 O—(CH₂)₃(thien-2,5-ylene)(CH₂)₃— —NHC(O)CH═CH— 25 O—(CH₂)₂C(O)NH(2-chloro-5-methoxyphen-1,4- —NHC(O)CH═CH— ylene)CH₂— 26 O—(CH₂)₇— —NHC(O)CH═CH— 27 O —(CH₂)₈— —NHC(O)CH═CH— 28 O—(CH₂)₂NHC(O)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 29 O1-[—(CH₂)₂C(O)](piperidin-4-yl)(CH₂)₂— —NHC(O)CH═CH— 30 O—(CH₂)₂NHC(O)(trans-cyclohex-1,4-ylene)CH₂— —NHC(O)CH═CH— 31 O—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)- —NHC(O)CH═CH— 32 O—(CH₂)₂NHC(O)NH(CH₂)₅— H —NHC(O)H 33 O —(CH₂)₂NH(phen-1,4-ylene)(CH₂)₂——NHC(O)CH═CH— 34 O —(CH₂)₃NHC(O)NH(CH₂)₅— H —NHC(O)H 35 O1-[—(CH₂)₂NHC(O)](piperidin-4-yl)(CH₂)₂— —NHC(O)CH═CH— 36 O—CH₂(phen-1,4-ylene)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 37 O—(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 38 NH—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 39 O—(CH₂)₂C(O)NHCH₂(pyrid-2,6-ylene)CH₂— —NHC(O)CH═CH— 40 O—(CH₂)₂C(O)NH(cis-cyclohex-1,4-ylene)CH₂— —NHC(O)CH═CH— 41 O—(CH₂)₂C(O)NH(trans-cyclohex-1,4-ylene)CH₂— —NHC(O)CH═CH— 42 O—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)CH₂— —NHC(O)CH═CH— 43 O—(CH₂)₂N(CH₃)C(O)(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 44 O—(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)CH₂— —NHC(O)CH═CH— 45 O—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂-(racemic) —NHC(O)CH═CH— 46 O—(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)— —NHC(O)CH═CH— ((S)-isomer) 47 O—(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)— —NHC(O)CH═CH— ((R)-isomer) 48 O2-[(S)—(—CH₂—](pyrrolidin-1-yl)C(O)(CH₂)₄— —NHC(O)CH═CH— 49 O2-[(S)—(—CH₂—](pyrrolidin-1-yl)C(O)(phen-1,4- —NHC(O)CH═CH— ylene)CH₂—50 O —(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— H —NHC(O)H 51 O—(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)— H —NHC(O)H ((R)-isomer) 52 O—(CH₂)₂C(O)NH(4-chlorophen-1,3-ylene)CH₂— —NHC(O)CH═CH— 53 NH—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 54 NH1-[—(CH₂)₂C(O)](piperidin-4-yl)(CH₂)₂— —NHC(O)CH═CH— 55 NH—(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 56 O—CH₂(2-fluorophen-1,3-ylene)CH₂— —NHC(O)CH═CH— 57 O—(CH₂)₂C(O)NH(4-methylphen-1,3-ylene)CH₂— —NHC(O)CH═CH— 58 O—(CH₂)₂C(O)NH(6-chlorophen-1,3-ylene)CH₂— —NHC(O)CH═CH— 59 O—(CH₂)₂C(O)NH(2,6-dichlorophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 60 O4-[—CH₂—](piperidin-1-yl)C(O)(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 61 O—(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂— H —NHC(O)H 62 O—(CH₂)₂C(O)N(CH₂CH₃)(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 63 O1-[—(CH₂)₂NHC(O)](piperidin-4-yl)- —NHC(O)CH═CH— 64 O—(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 65 O—(CH₂)₂NHC(O)(thien-2,5-ylene)CH₂— —NHC(O)CH═CH— 66 O—(CH₂)₂N(CH₃)C(O)(3-nitrophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 67 O—(CH₂)₂C(O)NH(trans-cyclohex-1,4-ylene)CH₂— H —NHC(O)H 68 O—(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)- —NHC(O)CH═CH— 69 O1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4- —NHC(O)CH═CH— yl)- 70 O5-[—(CH₂)₂NHC(O)](pyrid-2-yl)CH₂— —NHC(O)CH═CH— 71 O1-[—(CH₂)₃](piperidin-4-yl)CH₂— —NHC(O)CH═CH— 72 O—(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂— H —NHC(O)H 73 O—CH₂(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 74 O1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4- —NHC(O)CH═CH—yl)(CH₂)₂— 75 O —(CH₂)₃NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 76 O—(CH₂)₂C(O)NH(3-chlorophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 77 O—(CH₂)₂C(O)NH(2-(CF₃O—)phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 78 O—(CH₂)₃(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 79 O—CH₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 80 O—(CH₂)₂C(O)NH(2-iodophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 81 O—(CH₂)₂C(O)NH(2-chloro-6-methylphen-1,4- —NHC(O)CH═CH— ylene)CH₂— 82 O—(CH₂)₂C(O)NH(CH₂)₅-(racemic) —NHC(O)CH═CH— 83 O—(CH₂)₂C(O)NH(2-bromophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 84 O—(CH₂)₃(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 85 O1-[—CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4- —NHC(O)CH═CH—yl)(CH₂)₃— 86 O —(CH₂)₂C(O)NH(2-methoxyphen-1,4-ylene)CH₂— —NHC(O)CH═CH—87 O —(CH₂)₅NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 88 O4-[—(CH₂)₂—](piperidin-1-yl)(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 89 O—(CH₂)₂C(O)NH(phen-1,4-ylene)CH(CH₃)CH₂— —NHC(O)CH═CH— 90 O—(CH₂)₂-(trans-cyclohex-1,4-ylene)NH(phen-1,4- —NHC(O)CH═CH—ylene)(CH₂)₂— 91 O —(CH₂)₂C(O)NH(2-fluorophen-1,4-ylene)CH₂——NHC(O)CH═CH— 92 O —(CH₂)₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂——NHC(O)CH═CH— 93 O —(CH₂)₂C(O)NH(2,5-difluorophen-1,4-ylene)CH₂——NHC(O)CH═CH— 94 O —(CH₂)₂NHC(O)(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 95O 1-[—CH₂(pyrid-2,6-ylene)CH₂](piperidin-4-yl)CH₂— —NHC(O)CH═CH— 96 O—(CH₂)₂NH(naphth-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 97 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(phen-1,4- —NHC(O)CH═CH— ylene)CH₂— 98 O—(CH₂)₃(phen-1,4-ylene)NHC(O)(CH₂)₂— —NHC(O)CH═CH— 99 O—(CH₂)₃O(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 100 O2-[—(CH₂)₂](benzimidazol-5-yl)CH₂— —NHC(O)CH═CH— 101 O—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₂— —NHC(O)CH═CH— 102 O—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₄— —NHC(O)CH═CH— 103 O—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₅— —NHC(O)CH═CH— 104 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₂— —NHC(O)CH═CH— 105 O—(CH₂)₂NHC(O)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 106 O—(CH₂)₂N(CH₃)(CH₂)₂(cis-cyclohex-1,4-ylene)- —NHC(O)CH═CH— 107 O—(CH₂)₂C(O)NH(2,3,5,6-tetrafluorophen-1,4- —NHC(O)CH═CH— ylene)CH₂— 108O —(CH₂)₂C(O)NH(2,6-diiodophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 109 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₃— —NHC(O)CH═CH— 110 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₄— —NHC(O)CH═CH— 111 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₅— —NHC(O)CH═CH— 112 O—(CH₂)₂C(O)NHCH₂(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 113 O—(CH₂)₂C(O)NHCH₂(phen-1,4-ylene)CH₂— H —NHC(O)H 114 O—(CH₂)₂NHC(O)NHCH₂(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 115 O—(CH₂)₂NHC(O)NHCH₂(phen-1,4-ylene)CH₂— H —NHC(O)H 116 O—(CH₂)₂C(O)NH(2-methylphen-1,4-ylene)CH₂— —NHC(O)CH═CH— 117 O1-[—(CH₂)₃O(phen-1,4-ylene)(CH₂)₂](piperidin-4- —NHC(O)CH═CH— yl)CH₂—118 O —(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)(CH₂)₂— —NHC(O)CH═CH— 119 O—(CH₂)₂O(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 120 O—(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 121 O—(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 122 O—(CH₂)₂N(CH₃)C(O)(fur-2,5-ylene)CH₂— —NHC(O)CH═CH— 123 O—(CH₂)₂N(CH₃)C(O)(thien-2,5-ylene)CH₂— —NHC(O)CH═CH— 124 O—(CH₂)₂O(phen-1,4-ylene)O(CH₂)₂— —NHC(O)CH═CH— 125 O—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,4- —NHC(O)CH═CH—ylene)CH₂— 126 O —(CH₂)₂(trans-cyclohex-1,4- —NHC(O)CH═CH—ylene)NHC(O)CH₂O(phen-1,2-ylene)CH₂— 127 O —(CH₂)₂(trans-cyclohex-1,4-—NHC(O)CH═CH— ylene)NHC(O)CH₂O(phen-1,3-ylene)CH₂— 128 O—(CH₂)₂(trans-cyclohex-1,4- —NHC(O)CH═CH—ylene)NHC(O)CH₂O(phen-1,4-ylene)CH₂— 129 O—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(fur-2,5- —NHC(O)CH═CH—ylene)CH₂— 130 O —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(thien-—NHC(O)CH═CH— 2,5-ylene)CH₂— 131 O4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,2- —NHC(O)CH═CH— ylene)CH₂—132 O 4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,3- —NHC(O)CH═CH—ylene)CH₂— 133 O 4-[—(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,4-—NHC(O)CH═CH— ylene)CH₂— 134 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(fur-2,5-ylene)CH₂— —NHC(O)CH═CH— 135 O4-[—(CH₂)₂](piperidin-1-yl)C(O)(thien-2,5- —NHC(O)CH═CH— ylene)CH₂— 136O —(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,3- —NHC(O)CH═CH— ylene)CH₂— 137 O—(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,4- —NHC(O)CH═CH— ylene)CH₂— 138 O—(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,2- —NHC(O)CH═CH— ylene)CH₂— 139O —(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,3- —NHC(O)CH═CH— ylene)CH₂—140 O —(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,4- —NHC(O)CH═CH—ylene)CH₂— 141 O —(CH₂)₂(phen-1,4-ylene)NHC(O)(fur-2,5-ylene)CH₂——NHC(O)CH═CH— 142 O —(CH₂)₂(phen-1,4-ylene)NHC(O)(thien-2,5-—NHC(O)CH═CH— ylene)CH₂— 143 O—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,3- —NHC(O)CH═CH—ylene)CH₂— 144 O —(CH₂)₃O(phen-1,3-ylene)CH₂— —NHC(O)CH═CH— 145 O—CH₂CH(OH)CH₂NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 146 O—(CH₂)₄NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 147 O—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂NHC(O)CH₂— —NHC(O)CH═CH— 148 O—(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂NHC(O)CH₂— —NHC(O)CH═CH— 149 O—(CH₂)₂C(O)NHCH₂(trans-cyclohex-1,4-ylene)CH₂— —NHC(O)CH═CH— 150 O—(CH₂)₂NHC(O)(CH₂)₅— —NHC(O)CH═CH— 151 O—(CH₂)₂O(phen-1,3-ylene)O(CH₂)₂— —NHC(O)CH═CH— 152 O—(CH₂)₂O(phen-1,2-ylene)O(CH₂)₂— —NHC(O)CH═CH— 153 O—CH₂(phen-1,2-ylene)O(phen-1,2-ylene)CH₂— —NHC(O)CH═CH— 154 O—(CH₂)₂C(O)NH(CH₂)₆— —NHC(O)CH═CH— 155 O—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)- —NHC(O)CH═CH— 156 O—(CH₂)₃(phen-1,4-ylene)(CH₂)₃— —NHC(O)CH═CH— 157 O—(CH₂)₃(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 158 O—(CH₂)₄(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 159 O—(CH₂)₃(furan-2,5-ylene)(CH₂)₃— —NHC(O)CH═CH— 160 O—(CH₂)₂N(CH₃)C(O)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 161 O4-[—(CH₂)₂](piperidin-1-yl)C(O)NH(phen-1,4- —NHC(O)CH═CH— ylene)(CH₂)₂—162 O —(CH₂)₃(phen-1,3-ylene)(CH₂)₃— —NHC(O)CH═CH— 163 O—(CH₂)₃(tetrahydrofuran-2,5-ylene)(CH₂)₃— —NHC(O)CH═CH— 164 O—(CH₂)₂O(phen-1,4-ylene)C(O)(CH₂)₂— —NHC(O)CH═CH—¹In Tables I-III, “(racemic)” means the compound is racemic at thechiral carbon bearing the hydroxyl group in formula V, VI or VII.²For this group, the nitrogen atom is attached at R⁶ and carbon atom isattached at R⁷.

Another particular group of compounds of formula I are compounds offormula VI:

wherein W, R^(1A), R^(1B), R^(1C), R^(2A), R^(2B), R⁴, R⁶ and R⁷ are asdefined in Table II; or a pharmaceutically acceptable salt or solvatethereof. TABLE II Ex. W R^(1A) R^(1B) R^(1C) R^(2A) R^(2B) R⁴ R⁶ R⁷ 165O H H H Br H —(CH₂)₉— (racemic) —NHC(O)CH═CH— 166 O F H H H H —(CH₂)₉——NHC(O)CH═CH— 167 O H Cl H F F —(CH₂)₉— —NHC(O)CH═CH— 168 O H Cl Cl F F—(CH₂)₉— —NHC(O)CH═CH— 169 O H H H F F —(CH₂)₉— —NHC(O)CH═CH—

Another particular group of compounds of formula I are compounds offormula VII:

wherein W, R⁴, R⁶ and R⁷ are as defined in Table III; or apharmaceutically acceptable salt or solvate thereof. TABLE III Ex. W R⁴R⁶ R⁷ 170 O —(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 171 O—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂— H —NHC(O)H 172 O —(CH₂)₉——NHC(O)CH═CH— 173 O —(CH₂)₉— H —NHC(O)H 174 O —(CH₂)₂C(O)NH(CH₂)₅——NHC(O)CH₂CH₂— 175 O —(CH₂)₂C(O)NH(CH₂)₅— H —NHC(O)H 176 O—(CH₂)₂NHC(O)(CH₂)₅— —NHC(O)CH═CH— 177 O —(CH₂)₂NHC(O)(CH₂)₅— H —NHC(O)H178 O —(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂— —NHC(O)CH═CH— 179 O—(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂— H —NHC(O)H 180 O—(CH₂)₃(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂— —NHC(O)CH═CH— 181 O—(CH₂)₂NHC(O)(2-chlorophen-1,4-ylene)CH₂— —NHC(O)CH═CH— 182 O—(CH₂)₂NHC(O)(2-chloro-5-methoxyphen-1,4- —NHC(O)CH═CH— ylene)CH₂—

DEFINITIONS

When describing the compounds, compositions, methods and processes ofthis invention, the following terms have the following meanings unlessotherwise indicated.

The term “alkyl” means a monovalent saturated hydrocarbon group whichmay be linear or branched. Unless otherwise defined, such alkyl groupstypically contain from 1 to carbon atoms. Representative alkyl groupsinclude, by way of example, methyl, ethyl, n-propyl, isopropyl, n-butyl,sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,n-nonyl, n-decyl and the like.

The term “alkylene” means a divalent saturated hydrocarbon group whichmay be linear or branched. Unless otherwise defined, such alkylenegroups typically contain from 1 to 10 carbon atoms. Representativealkylene groups include, by way of example, methylene, ethane-1,2-diyl(“ethylene”), propane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl,pentane-1,5-diyl and the like.

The term “alkoxy” means a monovalent group of the formula (alkyl)-O—,where alkyl is as defined herein. Representative alkoxy groups include,by way of example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy,sec-butoxy, isobutoxy, tert-butoxy and the like.

The term “alkenyl” means a monovalent unsaturated hydrocarbon groupwhich may be linear or branched and which has at least one, andtypically 1, 2 or 3, carbon-carbon double bonds. Unless otherwisedefined, such alkenyl groups typically contain from 2 to 10 carbonatoms. Representative alkenyl groups include, by way of example,ethenyl, n-propenyl, isopropenyl, n-but-2-enyl, n-hex-3-enyl and thelike. The term “alkenylene” means a divalent alkenyl group.

The term “alkynyl” means a monovalent unsaturated hydrocarbon groupwhich may be linear or branched and which has at least one, andtypically 1, 2 or 3, carbon-carbon triple bonds. Unless otherwisedefined, such alkynyl groups typically contain from 2 to 10 carbonatoms. Representative alkynyl groups include, by way of example,ethynyl, n-propynyl, n-but-2-ynyl, n-hex-3-ynyl and the like. The term“alkynylene” means a divalent alkynyl group.

The term “aryl” means a monovalent aromatic hydrocarbon having a singlering (i.e., phenyl) or fused rings (i.e., naphthalene). Unless otherwisedefined, such aryl groups typically contain from 6 to 10 carbon ringatoms. Representative aryl groups include, by way of example, phenyl andnaphthalene-1-yl, naphthalene-2-yl, and the like. The term “arylene”means a divalent aryl group.

The term “azacycloalkyl” means a monovalent heterocyclic ring containingone nitrogen atom, i.e., a cycloalkyl group in which one carbon atom hasbeen replaced with a nitrogen atom. Unless otherwise defined, suchazacycloalkyl groups typically contain from 2 to 9 carbon atoms.Representative examples of an azacycloalkyl group are pyrrolidinyl andpiperidinyl groups. The term “azacycloalkylene” means a divalentazacycloakyl group. Representative examples of an azacycloalkylene groupare pyrrolidinylene and piperidinylene groups.

The term “cycloalkyl” means a monovalent saturated carbocyclichydrocarbon group. Unless otherwise defined, such cycloalkyl groupstypically contain from 3 to 10 carbon atoms. Representative cycloalkylgroups include, by way of example, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and the like. The term “cycloalkylene” means a divalentcycloalkyl group.

The term “halo” means fluoro, chloro, bromo and iodo.

The term “heteroaryl” means a monovalent aromatic group having a singlering or two fused rings and containing in the ring at least oneheteroatom (typically 1 to 3 heteroatoms) selected from nitrogen, oxygenor sulfur. Unless otherwise defined, such heteroaryl groups typicallycontain from 5 to 10 total ring atoms. Representative heteroaryl groupsinclude, by way of example, monovalent species of pyrrole, imidazole,thiazole, oxazole, furan, thiophene, triazole, pyrazole, isoxazole,isothiazole, pyridine, pyrazine, pyridazine, pyrimidine, triazine,indole, benzofuran, benzothiophene, benzimidazole, benzthiazole,quinoline, isoquinoline, quinazoline, quinoxaline and the like, wherethe point of attachment is at any available carbon or nitrogen ringatom. The term “heteroarylene” means a divalent heteroaryl group.

The term “heterocyclyl” or “heterocyclic” means a monovalent saturatedor unsaturated (non-aromatic) group having a single ring or multiplecondensed rings and containing in the ring at least one heteroatom(typically 1 to 3 heteroatoms) selected from nitrogen, oxygen or sulfur.Unless otherwise defined, such heterocyclic groups typically containfrom 2 to 9 total ring carbon atoms. Representative heterocyclic groupsinclude, by way of example, monovalent species of pyrrolidine,imidazolidine, pyrazolidine, piperidine, 1,4-dioxane, morpholine,thiomorpholine, piperazine, 3-pyrroline and the like, where the point ofattachment is at any available carbon or nitrogen ring atom. The term“heterocyclene” means a divalent heterocyclyl or heterocyclic group.

When a specific number of carbon atoms is intended for a particular termused herein, the number of carbon atoms is shown in parenthesespreceding the term. For example, the term “(1-4C)alkyl” means an alkylgroup having from 1 to 4 carbon atoms.

The term “pharmaceutically acceptable salt” means a salt which isacceptable for administration to a patient, such as a mammal (e.g.,salts having acceptable mammalian safety for a given dosage regime).Such salts can be derived from pharmaceutically acceptable inorganic ororganic bases and from pharmaceutically acceptable inorganic or organicacids. Salts derived from pharmaceutically acceptable inorganic basesinclude ammonium, calcium, copper, ferric, ferrous, lithium, magnesium,manganic, manganous, potassium, sodium, zinc and the like. Particularlypreferred are ammonium, calcium, magnesium, potassium and sodium salts.Salts derived from pharmaceutically acceptable organic bases includesalts of primary, secondary and tertiary amines, including substitutedamines, cyclic amines, naturally-occurring amines and the like, such asarginine, betaine, caffeine, choline, N,N′-dibenzylethylenediamine,diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol,ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine,glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperadine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. Salts derived frompharmaceutically acceptable acids include acetic, ascorbic,benzenesulfonic, benzoic, camphosulfonic, citric, ethanesulfonic,edisylic, fumaric, gentisic, gluconic, glucoronic, glutamic, hippuric,hydrobromic, hydrochloric, isethionic, lactic, lactobionic, maleic,malic, mandelic, methanesulfonic, mucic, naphthalenesulfonic,naphthalene-1,5-disulfonic, naphthalene-2,6-disulfonic, nicotinic,nitric, orotic, pamoic, pantothenic, phosphoric, succinic, sulfuric,tartaric, p-toluenesulfonic, xinafoic and the like. Particularlypreferred are citric, hydrobromic, hydrochloric, isethionic, maleic,naphthalene-1,5-disulfonic, phosphoric, sulfuric and tartaric acids.

The term “salt thereof” means a compound formed when the hydrogen of anacid is replaced by a cation, such as a metal cation or an organiccation and the like. Preferably, the salt is a pharmaceuticallyacceptable salt, although this is not required for salts of intermediatecompounds that are not intended for administration to a patient.

The term “solvate” means a complex or aggregate formed by one or moremolecules of a solute, i.e. a compound of formula I or apharmaceutically acceptable salt thereof, and one or more molecules of asolvent. Such solvates are typically crystalline solids having asubstantially fixed molar ratio of solute and solvent. Representativesolvents include, by way of example, water, methanol, ethanol,isopropanol, acetic acid and the like. When the solvent is water, thesolvate formed is a hydrate.

It will be appreciated that the term “or a pharmaceutically acceptablesalt or solvate or stereoisomer thereof” is intended to include allpermutations of salts, solvates and stereoisomers, such as a solvate ofa pharmaceutically acceptable salt of a stereoisomer of a compound offormula I.

The term “therapeutically effective amount” means an amount sufficientto effect treatment when administered to a patient in need of treatment.

The term “treating” or “treatment” as used herein means the treating ortreatment of a disease or medical condition (such as COPD) in a patient,such as a mammal (particularly a human) that includes:

-   -   (a) preventing the disease or medical condition from occurring,        i.e., prophylactic treatment of a patient;    -   (b) ameliorating the disease or medical condition, i.e.,        eliminating or causing regression of the disease or medical        condition in a patient;    -   (c) suppressing the disease or medical condition, i.e., slowing        or arresting the development of the disease or medical condition        in a patient; or    -   (d) alleviating the symptoms of the disease or medical condition        in a patient.

The term “leaving group” means a functional group or atom which can bedisplaced by another functional group or atom in a substitutionreaction, such as a nucleophilic substitution reaction. By way ofexample, representative leaving groups include chloro, bromo and iodogroups; sulfonic ester groups, such as mesylate, tosylate, brosylate,nosylate and the like; and acyloxy groups, such as acetoxy,trifluoroacetoxy and the like.

The term “protected derivatives thereof” means a derivative of thespecified compound in which one or more functional groups of thecompound are protected from undesired reactions with a protecting orblocking group. Functional groups which may be protected include, by wayof example, carboxylic acid groups, amino groups, hydroxyl groups, thiolgroups, carbonyl groups and the like. Representative protecting groupsfor carboxylic acids include esters (such as a p-methoxybenzyl ester),amides and hydrazides; for amino groups, carbamates (such astert-butoxycarbonyl) and amides; for hydroxyl groups, ethers and esters;for thiol groups, thioethers and thioesters; for carbonyl groups,acetals and ketals; and the like. Such protecting groups are well-knownto those skilled in the art and are described, for example, in T. W.Greene and G. M. Wuts, Protecting Groups in Organic Synthesis, ThirdEdition, Wiley, New York, 1999, and references cited therein.

The term “amino-protecting group” means a protecting group suitable forpreventing undesired reactions at an amino group. Representativeamino-protecting groups include, but are not limited to,tert-butoxycarbonyl (BOC), trityl (Tr), benzyloxycarbonyl (Cbz),9-fluorenylmethoxycarbonyl (Fmoc), formyl, trimethylsilyl (TMS),tert-butyldimethylsilyl (TBS), and the like.

The term “carboxy-protecting group” means a protecting group suitablefor preventing undesired reactions at a carboxy group. Representativecarboxy-protecting groups include, but are not limited to, esters, suchas methyl, ethyl, tert-butyl, benzyl (Bn), p-methoxybenzyl (PMB),9-fluoroenylmethyl (Fm), trimethylsilyl (TMS), tert-butyldimethylsilyl(TBS), diphenylmethyl (benzhydryl, DPM) and the like.

The term “hydroxyl-protecting group” means a protecting group suitablefor preventing undesirable reactions at a hydroxyl group. Representativehydroxyl-protecting groups include, but are not limited to, silyl groupsincluding tri(1-6C)alkylsilyl groups, such as trimethylsilyl (TMS),triethylsilyl (TES), tert-butyldimethylsilyl (TBS) and the like; esters(acyl groups) including (1-6C)alkanoyl groups, such as formyl, acetyland the like; arylmethyl groups, such as benzyl (Bn), p-methoxybenzyl(PMB), 9-fluorenylmethyl (Fm), diphenylmethyl (benzhydryl, DPM) and thelike. Additionally, two hydroxyl groups can also be protected as analkylidene group, such as prop-2-ylidine, formed, for example, byreaction with a ketone, such as acetone.

General Synthetic Procedures

The biphenyl derivatives of this invention can be prepared from readilyavailable starting materials using the following general methods andprocedures or by using other information readily available to those ofordinary skill in the art. Although a particular embodiment of thepresent invention may be shown or described herein, those skilled in theart will recognize that all embodiments or aspects of the presentinvention can be prepared using the methods described herein or by usingother methods, reagents and starting materials known to those skilled inthe art. It will also be appreciated that where typical or preferredprocess conditions (i.e., reaction temperatures, times, mole ratios ofreactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. While the optimumreaction conditions may vary depending on the particular reactants orsolvent used, such conditions can be readily determined by one skilledin the art by routine optimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary or desired to preventcertain functional groups from undergoing undesired reactions. Thechoice of a suitable protecting group for a particular functional groupas well as suitable conditions for protection and deprotection of suchfunctional groups are well-known in the art. Protecting groups otherthan those illustrated in the procedures described herein may be used,if desired. For example, numerous protecting groups, and theirintroduction and removal, are described in T. W. Greene and G. M. Wuts,Protecting Groups in Organic Synthesis, Third Edition, Wiley, New York,1999, and references cited therein.

By way of illustration, the biphenyl derivatives of this invention canbe prepared by a process comprising:

(a) reacting a compound of formula 1:

or a salt thereof, with a compound of formula 2:

wherein X¹ represents a leaving group, and P¹ and P² each independentlyrepresent a hydrogen atom or a hydroxyl-protecting group;

(b) reacting a compound of formula 3:

or salt thereof; with a compound of formula 4:

wherein X² represents a leaving group, and P³ and P⁴ each independentlyrepresent a hydrogen atom or a hydroxyl-protecting group;

(c) coupling a compound of formula 5:

with a compound of formula 6:

wherein X^(Qa) and X^(Qb) each independently represent functional groupsthat couple to form a group Q, P^(5a) represents a hydrogen atom or anamino-protecting group; and P^(5b) and P⁶ each independently represent ahydrogen atom or a hydroxyl-protecting group;

(d) for a compound of formula I wherein R⁵ represents a hydrogen atom,reacting a compound of formula 3 with a compound of formula 7:

or a hydrate thereof (e.g., a glyoxal), in the presence of a reducingagent, wherein P⁷ represents a hydrogen atom or a hydroxyl-protectinggroup;

(e) reacting a compound of formula 1 with a compound of formula 8:

or a hydrate thereof, in the presence of a reducing agent, wherein P⁸and P⁹ each independently represent a hydrogen atom or ahydroxyl-protecting group, P¹⁰ represents a hydrogen atom or anamino-protecting group, and R^(4′) represents a residue that, togetherwith the carbon to which it is attached, affords a group R⁴ uponcompletion of the reaction;

(f) reacting a compound of formula 9:

wherein X³ represents a leaving group, with a compound of formula 10:

wherein P¹¹ and P¹² each independently represent a hydrogen atom or ahydroxyl-protecting group, and P¹³ represents a hydrogen atom or anamino-protecting group; or

(g) reacting a compound of formula 11:

or a hydrate thereof; wherein R^(4′) represents a residue that, togetherwith the carbon to which it is attached, affords a group R⁴ uponcompletion of the reaction; with a compound of formula 10 in thepresence of a reducing agent; and then

removing any protecting group P¹, P², P³, P⁴, P^(5a), P^(5b), P⁶, P⁷,P⁸, P⁹, P¹⁰, P¹¹, P¹² or P¹³ to provide a compound of formula 1; andoptionally, forming a pharmaceutically acceptable salt thereof.

Generally, if a salt of one of the starting materials is used in theprocesses described above, such as an acid addition salt, the salt istypically neutralized before or during the reaction process. Thisneutralization reaction is typically accomplished by contacting the saltwith one molar equivalent of a base for each molar equivalent of acidaddition salt.

In process (a), i.e., the reaction between the compounds of formula 1and 2, the leaving group represented by X¹ can be, for example, halo,such as chloro, bromo or iodo, or a sulfonic ester group, such asmesylate or tosylate. The groups P¹ and P² can be, for example,trimethylsilyl and benzyl, respectively. This reaction is typicallyconducted in an inert diluent, such as acetonitrile, in the presence ofa base. For example, this reaction can be conducted in the presence of atertiary amine, such as diisopropylethylamine. Generally, this reactionis conducted at a temperature in the range of from 0° C. to 100° C.until the reaction is substantially complete. The reaction product isthen isolated using conventional procedures, such as extraction,recrystallization, chromatography and the like.

Compounds of formula 1 are generally known in the art or can be preparedfrom commercially available starting materials and reagents usingwell-known procedures. For example, compounds of formula 1 can beprepared by deprotecting a compound of formula 12:

wherein P¹⁴ represents an amino-protecting group, such as a benzylgroup. By way of illustration, a benzyl group can be readily removed byreduction using, for example, hydrogen or ammonium formate and a groupVIII metal catalyst, such as palladium on carbon. When W representsNW^(a), the hydrogenation reaction is conveniently performed usingPearlman's catalyst (i.e., Pd(OH)₂).

Compounds of formula 12 can be prepared by reacting an isocyanatecompound of formula 13:

with a compound of formula 14:

Compounds of formula 2 can be prepared by various procedures describedherein or by procedures that are well-known to those skilled in the art.For example, the hydroxyl group of a compound of formula 23 below, canbe readily converted into a leaving group using well-known reagents andprocedures. By way of illustration, a hydroxyl group can be convertedinto a halo group using an inorganic acid halide, such as thionylchloride, phosphorous trichloride, phosphorous tribromide, phosphorousoxychloride and the like, or a halogen acid, such a hydrogen bromide.

In process (b), i.e., the reaction of a compound of formula 3 with acompound of formula 4, the leaving represented by X² can be, forexample, halo, such as chloro, bromo or iodo, or a sulfonic ester group,such as mesylate or tosylate. The groups P³ and P⁴ can be, for example,tert-butyldimethylsilyl and benzyl, respectively. This reaction istypically conducted in the presence of a base, such as sodiumbicarbonate, and an alkali metal iodide, such as sodium iodide.Generally, this reaction is conducted in an inert diluent, such astetrahydrofuran, at a temperature ranging from 25° C. to 100° C. untilthe reaction is substantially complete. The reaction product is thenisolated using conventional procedures, such as extraction,recrystallization, chromatography and the like.

Compounds of formula 3 can be prepared by deprotecting a compound offormula 15:

wherein one or both of P¹⁵ and P¹⁶ independently represents a protectinggroup, such as tert-butoxycarbonyl, and any remainder represents ahydrogen atom. For example, a tert-butoxycarbonyl group can be removedby treating the protected compound with trifluoroacetic acid.

Compounds of formula 15 can be prepared by reacting a compound offormula 1 with a compound of formula 16:X³—R⁴—NP¹⁵P¹⁶  16

wherein X³ represents a leaving group such as halo, such as chloro,bromo or iodo, or sulfonic ester group, such as mesylate or tosylate.This reaction is typically conducted by contacting a compound of formula1 with a compound of formula 16 in an inert diluent, such asacetonitrile, DMF or mixtures thereof, at a temperature ranging fromabout 0° C. to about 100° C. until the reaction is substantiallycomplete.

Alternatively, compounds of formula 3 can be obtained by reductiveamination of a compound of formula 11. The reductive amination can beperformed by reacting the compound of formula 11 with, for example,benzylamine and hydrogen in the presence of palladium on carbon.

Compounds of formula 11 may be prepared by oxidizing the correspondingalcohol of formula 17:

using a suitable oxidizing agent, such as sulfur trioxide pyridinecomplex and dimethyl sulfoxide. This oxidation reaction is typicallyconducted in an inert diluent, such as dichloromethane, the presence ofa tertiary amine, such as diisopropylethylamine, at a temperatureranging from about −20° C. to about 25° C.

Compounds of formula 17 can be prepared by reacting a compound offormula 1 with a compound of formula 18:X⁴—R⁴—OH  18wherein X⁴ represents a leaving group such as halo, such as chloro,bromo or iodo, or a sulfonic ester group, such as mesylate or tosylate.

Compounds of formula 4 can be prepared by reacting a compound of formula19:

with a reducing agent, such as borane. If desired, such a reduction canbe performed in the presence of a chiral catalyst to provide compoundsof formula 4 in chiral form. For example, compounds of formula 19 can bereduced in the presence of a chiral catalyst formed from(R)-(+)-α,α-diphenyl-2-pyrrolidinemethanol and trimethylboroxine; oralternatively, from (S)-(−)-α,α-diphenyl-2-pyrrolidinemethanol andtrimethylboroxine. The resulting hydroxyl group can then be protectedwith a hydroxyl-protecting group, P³, by reaction with, for example,tert-butyldimethylsilyl trifluoromethanesulfonate.

Compounds of formula 19 in which X² represents a bromine atom can beprepared by reacting a compound of formula 20:

with bromine in the presence of a Lewis acid, such as boron trifluoridediethyl etherate. Compounds of formula 20 are well-known in the art orcan be prepared by well-known procedures using commercially availablestarting materials and reagents.

Referring to process (c), i.e., the reaction of a compound of formula 5with a compound of formula 6, it will be appreciated that the groupsX^(Qa) and X^(Qb) should be selected so as to afford the desired group Qupon completion of the reaction. For example, when the desired group Qis an amide group, i.e., —N(Q^(a))C(O)— or —C(O)N(Q^(b)), one of X^(Qa)and X^(Qb) can be an amine group (i.e., —NHQ^(a) or —NHQ^(b)) and theother can be a carboxyl group (i.e., —COOH) or a reactive derivativethereof (such as acyl halide, such as an acyl chloride or acyl bromide).The groups P^(5a), P^(5b) and P⁶ can be, for example, benzyl,trimethylsilyl and benzyl, respectively. When Q is an amide group, thereaction can be performed under conventional amide coupling conditions.Similarly, when the desired group Q is a sulfonamide, i.e.,—N(Q^(c))S(O)₂— or —S(O)₂N(Q^(d))-, one of X^(Qa) and X^(Qb) can be anamine group, —NHQ^(c) or —NHQ^(d) and the other can be a sulfonyl halidegroup (such as sulfonyl chloride or sulfonyl bromide).

Compounds of formula 5 can be prepared by reacting a compound of formula1 with a compound of formula 21:X⁵—(R^(4a))_(d)-(A¹)_(e)-(R^(4b))_(f)—X^(Qa′)  21wherein X⁵ represents a leaving group including halo, such as chloro,bromo or iodo, and a sulfonic ester group, such as mesylate or tosylate;and X^(Qa′) represents X^(Qa), such as a carboxyl group or an aminogroup NHQ^(a), or a protected derivative thereof, such as a(1-6C)alkoxycarbonylamino group or a tert-butoxycarbonylamino group.This reaction is typically conducted by a method analogous to that usedto prepare compounds of formula 3, followed by removing any protectinggroup in X^(Qa′).

Compounds of formula 6 can be prepared by reacting a compound of formula4 with a compound of formula 22:X^(Qb′)—(R^(4c))_(g)-(A²)_(h)-(R^(4d))_(i)—X⁶  22wherein X⁶ represents a leaving group including halo, such as chloro,bromo or iodo, and a sulfonic ester group, such as mesylate or tosylate;and X^(Qb′) represents X^(Qb), such as a carboxyl group or an aminogroup NHQ^(b), or a protected derivative thereof, such as a(1-6C)alkoxycarbonyl group or a tert-butoxycarbonylamino group. Thisreaction is typically conducted by a method analogous to that used toprepare compounds of formula 3, followed by removing any protectinggroup in X^(Qb′).

Referring to process (d), i.e., the reaction of a compound of formula 3with a compound of formula 7, any suitable reducing agent may be used inthis reaction. For example, the reducing agent can be hydrogen in thepresence of a Group VIII metal catalyst, such as palladium on carbon; ora metal hydride reagent, such as sodium triacetoxyborohydride. The groupP⁷ can be, for example, benzyl. This reaction is typically conducted inan inert diluent and a protic solvent, such as a mixture ofdichloroethane and methanol, at a temperature in the range of from 0° C.to 100° C. until the reaction is substantially complete.

Compounds of formula 7 in the form of a hydrate can be prepared byconventional procedures, for example, by dibrominating a compound offormula 19 (where X² in this case can also be hydrogen), and thenhydrolyzing the resulting dibromide to form a glyoxal or a hydratethereof. For example, a compound of formula 19 can be reacted withhydrogen bromide and then hydrolyzed with water to form thecorresponding glyoxal hydrate.

Referring to process (e), i.e., the reaction of a compound of formula 1with a compound of formula 8, any suitable reducing agent may be used inthis reaction. For example, the reducing agent may be hydrogen in thepresence of a Group VIII metal catalyst, such as palladium on carbon; ora metal hydride reagent, such as sodium triacetoxyborohydride. Thegroups P⁸, P⁹ and P¹⁰ can be, for example, trimethylsilyl, benzyl andbenzyl, respectively. Typically, this reduction reaction is conducted inan inert diluent and a protic solvent, such as dichloroethane andmethanol, at a temperature in the range of from 0° C. to 100° C. untilthe reaction is substantially complete.

Compounds of formula 8 may be prepared by oxidizing a compound offormula 23:

using any suitable oxidizing agent, such as sulfur trioxide pyridinecomplex and dimethyl sulfoxide. This reaction is typically conducted inthe presence of a tertiary amine, such as diisopropylethylamine, at atemperature in the range of from about −20° C. to about 25° C. until theoxidation is substantially complete.

Compounds of formula 23 can be prepared by reacting a compound offormula 10 with a compound of formula 24:HO—R⁴—X⁷  24wherein X⁷ represents a leaving group including halo, such as chloro,bromo or iodo, and a sulfonic ester group, such as mesylate or tosylate.

Referring to process (f), i.e., the reaction of a compound of formula 9with a compound of formula 10, the leaving group represented by X³ canbe, for example, halo, such as chloro, bromo or iodo, or a sulfonicester group, such as mesylate or tosylate. The groups P¹¹, P¹² and P¹³can be, for example, trimethylsilyl, benzyl and benzyl, respectively.This reaction is typically conducted an inert diluent, such asacetonitrile, in the presence of a suitable base. For example, thisreaction can be conducted in the presence of a tertiary amine, such asdiisopropylethylamine. Generally, this reaction is conducted at atemperature in the range of from 0° C. to 100° C. until the reaction issubstantially complete.

Compounds of formula 9 can be prepared by steps analogous to those ofmethods (a) to (e) herein, starting from a compound of formula 1.Additionally, compounds of formula 10 can be prepared from compounds offormula 4 by reaction with an amine of formula P¹³NH₂.

Referring to process (g), i.e., the reaction of a compound of formula IIwith a compound of formula 10, any suitable reducing agent may be usedin this reaction. For example, the reducing agent may be hydrogen in thepresence of a Group VIII metal catalyst, such as palladium on carbon; ora metal hydride reagent, such as sodium triacetoxyborohydride. Thegroups P¹¹, P¹² and P¹³ can be, for example, tert-butyldimethylsilyl,benzyl and benzyl, respectively. Typically, this reduction reaction isconducted in an inert diluent and a protic solvent, such asdichloroethane and methanol, at a temperature in the range of from 0° C.to 100° C. until the reaction is substantially complete.

Compounds of formula 11 are readily prepared by oxidation of thecorresponding alcohol or by hydrolysis of the corresponding acetal. Anysuitable oxidizing agent may be employed in this reaction to provide thealdehyde, such as sulfur trioxide pyridine complex and dimethylsulfoxide. The acetal may be hydrolyzed under conventional conditionsusing aqueous acid to provide the aldehyde.

In a particular embodiment, certain compounds of formula I are preparedby a process comprising:

(h) deprotecting a compound of formula 25:

wherein P¹⁷ represents a hydrogen atom or an amino-protecting group; andeach of P¹⁸, P¹⁹ and P²⁰ independently represent a hydrogen atom or ahydroxyl-protecting group; provided that at least one of P¹⁷, P¹⁸, P¹⁹or P²⁰ is a protecting group;

(i) deprotecting a compound of formula 26:

wherein P²¹ represents a hydrogen atom or an amino-protecting group; andeach of P²² and P²³ independently represent a hydrogen atom or ahydroxyl-protecting group; provided that at least one of P²¹, P²² or P²³is a protecting group; or

(j) deprotecting a compound of formula 27:

wherein P²⁴ represents a hydrogen atom or an amino-protecting group; andeach of P²⁵ and P²⁶ independently represent a hydrogen atom or ahydroxyl-protecting group; provided that at least one of P²⁴, P²⁵ or P²⁶is a protecting group.

to provide a compound of formula I and, optionally, forming apharmaceutically acceptable salt of the compound of formula I.

Referring to process (h), examples of particular values for P¹⁷, P¹⁸,P¹⁹ and P²⁰ are: for P¹⁷, hydrogen or benzyl; for P¹⁸ hydrogen ortert-butyldimethylsilyl; and for P¹⁹ and P²⁰ hydrogen or benzyl, ortogether propylidine. In this process, benzyl protecting groups areconveniently removed by catalytic hydrogenation in the presence of aGroup VIII metal catalyst, such as palladium on carbon; atert-butyldimethylsilyl group is conveniently removed by treatment withhydrogen fluoride, such as triethylamine trihydrofluoride; and apropylidine group is conveniently removed by treatment with an acid,such as trifluoroacetic acid.

Compounds of formula 25 can be prepared by the methods described herein,such as by processes (a) to (g). Alternatively, compounds of formula 25can be prepared by reacting a compound of formula 28:

wherein R⁸ represents —CH₂OP¹⁹, —CHO, —COOH or —C(O)O(1-6C)alkoxy, suchas carbomethoxy, R⁹ represents OP¹⁸ and R¹⁰ represents a hydrogen atom,or R⁹ and R¹⁰ together represent ═O, with a reducing agent. Any suitablereducing agent may be used in this reaction including, by way ofexample, metal hydride reducing agents, such as sodium borohydride,lithium aluminum hydride and the like.

Compounds of formula 28 in which R⁹ and R¹⁰ together represent a ═Ogroup can be readily prepared by reacting a compound of formula 29:

or a salt thereof, with a compound of formula 30:

wherein X⁸ represents a leaving group, such as a bromo.

Referring to process (i), examples of particular values for P²¹, P²² andP²³ are: for P²¹, hydrogen or benzyl; for P²² hydrogen ortert-butyldimethylsilyl; and for P²³ hydrogen or benzyl. In thisprocess, benzyl protecting groups are conveniently removed by catalytichydrogenation in the presence of a Group VIII metal catalyst, such aspalladium on carbon; and a tert-butyldimethylsilyl group is convenientlyremoved by treatment with hydrogen fluoride, such as triethylaminetrihydrofluoride. Compounds of formula 26 can be prepared by the methodsdescribed herein, such as by processes (a) to (g).

Referring to process (O), examples of particular values for P²⁴, P²⁵ andP²⁶ are: for P²⁴, hydrogen or benzyl; for P²⁵ hydrogen ortert-butyldimethylsilyl; and for P²⁶ hydrogen or benzyl. In thisprocess, benzyl protecting groups are conveniently removed by catalytichydrogenation in the presence of a Group VIII metal catalyst, such aspalladium on carbon; and a tert-butyldimethylsilyl group is convenientlyremoved by treatment with hydrogen fluoride, such as triethylaminetrihydrofluoride. Compounds of formula 27 can be prepared by the methodsdescribed herein, such as by processes (a) to (g).

Additionally, compounds of formula I in which R⁶ and R⁷ together form—NR^(7g)C(O)—CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)—NR^(7p)— may be prepared by reducing acorresponding compound of formula I in which R⁶ and R⁷ together form—NR^(7a)C(O)—CR^(7b)═CR^(7c) or —CR^(7d)═CR^(7e)—C(O)—NR^(7f)—, forexample by catalytic hydrogenation as described in Example 6hereinafter.

Further details regarding specific reaction conditions and otherprocedures for preparing representative compounds of this invention orintermediates thereof are described in the Examples set forth below.

Pharmaceutical Compositions and Formulations

The biphenyl derivatives of this invention are typically administered toa patient in the form of a pharmaceutical composition or formulation.Such pharmaceutical compositions may be administered to the patient byany acceptable route of administration including, but not limited to,inhaled, oral, nasal, topical (including transdermal) and parenteralmodes of administration. It will be understood that any form of thecompounds of this invention, (i.e., free base, pharmaceuticallyacceptable salt, solvate, etc.) that is suitable for the particular modeof administration can be used in the pharmaceutical compositionsdiscussed herein.

Accordingly, in one of its compositions aspects, this invention isdirected to a pharmaceutical composition comprising a pharmaceuticallyacceptable carrier or excipient and a therapeutically effective amountof a compound of formula I, or a pharmaceutically acceptable saltthereof. Optionally, such pharmaceutical compositions may contain othertherapeutic and/or formulating agents if desired.

The pharmaceutical compositions of this invention typically contain atherapeutically effective amount of a compound of the present inventionor a pharmaceutically acceptable salt thereof. Typically, suchpharmaceutical compositions will contain from about 0.01 to about 95% byweight of the active agent; including, from about 0.01 to about 30% byweight; such as from about 0.01 to about 10% by weight of the activeagent.

Any conventional carrier or excipient may be used in the pharmaceuticalcompositions of this invention. The choice of a particular carrier orexcipient, or combinations of carriers or exipients, will depend on themode of administration being used to treat a particular patient or typeof medical condition or disease state. In this regard, the preparationof a suitable pharmaceutical composition for a particular mode ofadministration is well within the scope of those skilled in thepharmaceutical arts. Additionally, the ingredients for such compositionsare commercially available from, for example, Sigma, P.O. Box 14508, St.Louis, Mo. 63178. By way of further illustration, conventionalformulation techniques are described in Remington: The Science andPractice of Pharmacy, 20^(th) Edition, Lippincott Williams & White,Baltimore, Md. (2000); and H. C. Ansel et al., Pharmaceutical DosageForms and Drug Delivery Systems, 7^(th) Edition, Lippincott Williams &White, Baltimore, Md. (1999).

Representative examples of materials which can serve as pharmaceuticallyacceptable carriers include, but are not limited to, the following: (1)sugars, such as lactose, glucose and sucrose; (2) starches, such as cornstarch and potato starch; (3) cellulose, and its derivatives, such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;(4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8)excipients, such as cocoa butter and suppository waxes; (9) oils, suchas peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil,corn oil and soybean oil; (10) glycols, such as propylene glycol; (I 1)polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol;(12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14)buffering agents, such as magnesium hydroxide and aluminum hydroxide;(15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18)Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions;(21) compressed propellant gases, such as chlorofluorocarbons andhydrofluorocarbons; and (22) other non-toxic compatible substancesemployed in pharmaceutical compositions.

The pharmaceutical compositions of this invention are typically preparedby thoroughly and intimately mixing or blending a compound of theinvention with a pharmaceutically acceptable carrier and one or moreoptional ingredients. If necessary or desired, the resulting uniformlyblended mixture can then be shaped or loaded into tablets, capsules,pills, canisters, cartridges, dispensers and the like using conventionalprocedures and equipment.

In one embodiment, the pharmaceutical compositions of this invention aresuitable for inhaled administration. Suitable pharmaceuticalcompositions for inhaled administration will typically be in the form ofan aerosol or a powder. Such compositions are generally administeredusing well-known delivery devices, such as a nebulizer inhaler, ametered-dose inhaler (MDI), a dry powder inhaler (DPI) or a similardelivery device.

In a specific embodiment of this invention, the pharmaceuticalcomposition comprising the active agent is administered by inhalationusing a nebulizer inhaler. Such nebulizer devices typically produce astream of high velocity air that causes the pharmaceutical compositioncomprising the active agent to spray as a mist that is carried into thepatient's respiratory tract. Accordingly, when formulated for use in anebulizer inhaler, the active agent is typically dissolved in a suitablecarrier to form a solution. Alternatively, the active agent can bemicronized and combined with a suitable carrier to form a suspension ofmicronized particles of respirable size, where micronized is typicallydefined as having about 90% or more of the particles with a diameter ofless than about 10 μm. Suitable nebulizer devices are providedcommercially, for example, by PARI GmbH (Starnberg, German). Othernebulizer devices include Respimat (Boehringer Ingelheim) and thosedisclosed, for example, in U.S. Pat. No. 6,123,068 and WO 97/12687.

A representative pharmaceutical composition for use in a nebulizerinhaler comprises an isotonic aqueous solution comprising from about0.05 μg/mL to about 10 mg/mL of a compound of formula I or apharmaceutically acceptable salt or solvate or stereoisomer thereof.

In another specific embodiment of this invention, the pharmaceuticalcomposition comprising the active agent is administered by inhalationusing a dry powder inhaler. Such dry powder inhalers typicallyadminister the active agent as a free-flowing powder that is dispersedin a patient's air-stream during inspiration. In order to achieve a freeflowing powder, the active agent is typically formulated with a suitableexcipient such as lactose or starch.

A representative pharmaceutical composition for use in a dry powderinhaler comprises dry lactose having a particle size between about 1 μmand about 100 μm and micronized particles of a compound of formula I, ora pharmaceutically acceptable salt or solvate or stereoisomer thereof.

Such a dry powder formulation can be made, for example, by combining thelactose with the active agent and then dry blending the components.Alternatively, if desired, the active agent can be formulated without anexcipient. The pharmaceutical composition is then typically loaded intoa dry powder dispenser, or into inhalation cartridges or capsules foruse with a dry powder delivery device.

Examples of dry powder inhaler delivery devices include Diskhaler(GlaxoSmithKline, Research Triangle Park, N.C.) (see, e.g., U.S. Pat.No. 5,035,237); Diskus (GlaxoSmithKline) (see, e.g., U.S. Pat. No.6,378,519; Turbuhaler (AstraZeneca, Wilmington, Del.) (see, e.g., U.S.Pat. No. 4,524,769); Rotahaler (GlaxoSmithKline) (see, e.g., U.S. Pat.No. 4,353,365) and Handihaler (Boehringer Ingelheim). Further examplesof suitable DPI devices are described in U.S. Pat. Nos. 5,415,162,5,239,993, and 5,715,810 and references cited therein.

In yet another specific embodiment of this invention, the pharmaceuticalcomposition comprising the active agent is administered by inhalationusing a metered-dose inhaler. Such metered-dose inhalers typicallydischarge a measured amount of the active agent or a pharmaceuticallyacceptable salt thereof using compressed propellant gas. Accordingly,pharmaceutical compositions administered using a metered-dose inhalertypically comprise a solution or suspension of the active agent in aliquefied propellant. Any suitable liquefied propellant may be employedincluding chlorofluorocarbons, such as CCl₃F, and hydrofluoroalkanes(HFAs), such as 1,1,1,2-tetrafluoroethane (HFA 134a) and1,1,1,2,3,3,3-heptafluoro-n-propane, (HFA 227). Due to concerns aboutchlorofluorocarbons affecting the ozone layer, formulations containingHFAs are generally preferred. Additional optional components of HFAformulations include co-solvents, such as ethanol or pentane, andsurfactants, such as sorbitan trioleate, oleic acid, lecithin, andglycerin. See; for example, U.S. Pat. No. 5,225,183, EP 0717987 A2, andWO 92/22286.

A representative pharmaceutical composition for use in a metered-doseinhaler comprises from about 0.01% to about 5% by weight of a compoundof formula I, or a pharmaceutically acceptable salt or solvate orstereoisomer thereof; from about 0% to about 20% by weight ethanol; andfrom about 0% to about 5% by weight surfactant; with the remainder beingan HFA propellant.

Such compositions are typically prepared by adding chilled orpressurized hydrofluoroalkane to a suitable container containing theactive agent, ethanol (if present) and the surfactant (if present). Toprepare a suspension, the active agent is micronized and then combinedwith the propellant. The formulation is then loaded into an aerosolcanister, which forms a portion of a metered-dose inhaler device.Examples of metered-dose inhaler devices developed specifically for usewith HFA propellants are provided in U.S. Pat. Nos. 6,006,745 and6,143,277. Alternatively, a suspension formulation can be prepared byspray drying a coating of surfactant on micronized particles of theactive agent. See, for example, WO 99/53901 and WO 00/61108.

For additional examples of processes of preparing respirable particles,and formulations and devices suitable for inhalation dosing see U.S.Pat. Nos. 6,268,533, 5,983,956, 5,874,063, and 6,221,398, and WO99/55319 and WO 00/30614.

In another embodiment, the pharmaceutical compositions of this inventionare suitable for oral administration. Suitable pharmaceuticalcompositions for oral administration may be in the form of capsules,tablets, pills, lozenges, cachets, dragees, powders, granules; or as asolution or a suspension in an aqueous or non-aqueous liquid; or as anoil-in-water or water-in-oil liquid emulsion; or as an elixir or syrup;and the like; each containing a predetermined amount of a compound ofthe present invention as an active ingredient.

When intended for oral administration in a solid dosage form (i.e., ascapsules, tablets, pills and the like), the pharmaceutical compositionsof this invention will typically comprise a compound of the presentinvention as the active ingredient and one or more pharmaceuticallyacceptable carriers, such as sodium citrate or dicalcium phosphate.Optionally or alternatively, such solid dosage forms may also comprise:(1) fillers or extenders, such as starches, lactose, sucrose, glucose,mannitol, and/or silicic acid; (2) binders, such ascarboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,sucrose and/or acacia; (3) humectants, such as glycerol; (4)disintegrating agents, such as agar-agar, calcium carbonate, potato ortapioca starch, alginic acid, certain silicates, and/or sodiumcarbonate; (5) solution retarding agents, such as paraffin; (6)absorption accelerators, such as quaternary ammonium compounds; (7)wetting agents, such as cetyl alcohol and/or glycerol monostearate; (8)absorbents, such as kaolin and/or bentonite clay; (9) lubricants, suchas talc, calcium stearate, magnesium stearate, solid polyethyleneglycols, sodium lauryl sulfate, and/or mixtures thereof; (10) coloringagents; and (11) buffering agents.

Release agents, wetting agents, coating agents, sweetening, flavoringand perfuming agents, preservatives and antioxidants can also be presentin the pharmaceutical compositions of this invention. Examples ofpharmaceutically acceptable antioxidants include: (1) water-solubleantioxidants, such as ascorbic acid, cysteine hydrochloride, sodiumbisulfate, sodium metabisulfate sodium sulfite and the like; (2)oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and (3) metal-chelating agents,such as citric acid, ethylencdiamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like. Coating agents fortablets, capsules, pills and like, include those used for entericcoatings, such as cellulose acetate phthalate (CAP), polyvinyl acetatephthalate (PVAP), hydroxypropyl methylcellulose phthalate, methacrylicacid-methacrylic acid ester copolymers, cellulose acetate trimellitate(CAT), carboxymethyl ethyl cellulose (CMEC), hydroxypropyl methylcellulose acetate succinate (HPMCAS), and the like.

If desired, the pharmaceutical compositions of the present invention mayalso be formulated to provide slow or controlled release of the activeingredient using, by way of example, hydroxypropyl methyl cellulose invarying proportions; or other polymer matrices, liposomes and/ormicrospheres.

In addition, the pharmaceutical compositions of the present inventionmay optionally contain opacifying agents and may be formulated so thatthey release the active ingredient only, or preferentially, in a certainportion of the gastrointestinal tract, optionally, in a delayed manner.Examples of embedding compositions which can be used include polymericsubstances and waxes. The active ingredient can also be inmicro-encapsulated form, if appropriate, with one or more of theabove-described excipients.

Suitable liquid dosage forms for oral administration include, by way ofillustration, pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. Such liquid dosage formstypically comprise the active ingredient and an inert diluent, such as,for example, water or other solvents, solubilizing agents andemulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate,ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol,1,3-butylene glycol, oils (esp., cottonseed, groundnut, corn, germ,olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol,polyethylene glycols and fatty acid esters of sorbitan, and mixturesthereof. Suspensions, in addition to the active ingredient, may containsuspending agents such as, for example, ethoxylated isostearyl alcohols,polyoxyethylene sorbitol and sorbitan esters, microcrystallinecellulose, aluminium metahydroxide, bentonite, agar-agar and tragacanth,and mixtures thereof.

When intended for oral administration, the pharmaceutical compositionsof this invention are preferably packaged in a unit dosage form. Theterm “unit dosage form” means a physically discrete unit suitable fordosing a patient, i.e., each unit containing a predetermined quantity ofactive agent calculated to produce the desired therapeutic effect eitheralone or in combination with one or more additional units. For example,such unit dosage forms may be capsules, tablets, pills, and the like.

The compounds of this invention can also be administered transdermallyusing known transdermal delivery systems and excipents. For example, acompound of this invention can be admixed with permeation enhancers,such as propylene glycol, polyethylene glycolm monolaurate,azacycloalkan-2-ones and the like, and incorporated into a patch orsimilar delivery system. Additional excipients including gelling agents,emulsifiers and buffers, may be used in such transdermal compositions ifdesired.

The pharmaceutical compositions of this invention may also contain othertherapeutic agents that are co-administered with a compound of formulaI, or pharmaceutically acceptable salt or solvate or stereoisomerthereof. For example, the pharmaceutical compositions of this inventionmay further comprise one or more therapeutic agents selected from otherbronchodilators (e.g., PDE₃ inhibitors, adenosine 2b modulators and β₂adrenergic receptor agonists); anti-inflammatory agents (e.g. steroidalanti-inflammatory agents, such as corticosteroids; non-steroidalanti-inflammatory agents (NSAIDs), and PDE₄ inhibitors); othermuscarinic receptor antagonists (i.e., antichlolinergic agents);antiinfective agents (e.g. Gram positive and Gram negative antibioticsor antivirals); antihistamines; protease inhibitors; and afferentblockers (e.g., D₂ agonists and neurokinin modulators). The othertherapeutic agents can be used in the form of pharmaceuticallyacceptable salts or solvates. Additionally, if appropriate, the othertherapeutic agents can be used as optically pure stereoisomers.

Representative β₂ adrenergic receptor agonists that can be used incombination with, and in addition to, the compounds of this inventioninclude, but are not limited to, salmeterol, salbutamol, formoterol,salmefamol, fenoterol, terbutaline, albuterol, isoetharine,metaproterenol, bitolterol, pirbuterol, levalbuterol and the like, orpharmaceutically acceptable salts thereof. Other β₂ adrenergic receptoragonists that can be used in combination with the compounds of thisinvention include, but are not limited to,3-(4-{[6-({(2R)-2-hydroxy-2-[4-hydroxy-3-(hydroxymethyl)-phenyl]ethyl}amino)-hexyl]oxy}butyl)benzenesulfonamideand3-(-3-{[7-({(2R)-2-hydroxy-2-[4-hydroxy-3-(hydroxymethyl)phenyl]ethyl}-amino)heptyl]oxy}-propyl)benzenesulfonamideand related compounds disclosed in WO 02/066422, published on Aug. 29,2002;3-[3-(4-{[6-([(2R)-2-hydroxy-2-[4-hydroxy-3-(hydroxymethyl)phenyl]ethyl}amino)hexyl]oxy}butyl)-phenyl]imidazolidine-2,4-dioneand related compounds disclosed in WO 02/070490, published Sep. 12,2002;3-(4-{[6-({(2R)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)hexyl]oxy}butyl)-benzenesulfonamide,3-(4-{[6-({(2S)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)hexyl]oxy}butyl)-benzenesulfonamide,3-(4-{[6-({(2R/S)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)hexyl]oxy}butyl)-benzenesulfonamide,N-(tert-butyl)-3-(4-{[6-({(2R)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)hexyl]-oxy}butyl)benzenesulfonamide,N-(tert-butyl)-3-(4-{[6-({(2S)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)-hexyl]oxy}butyl)-benzenesulfonamide,N-(tert-butyl)-3-(4-{[6-({(2R/S)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)hexyl]-oxy}butyl)benzenesulfonamideand related compounds disclosed in WO 02/076933, published on Oct. 3,2002;4-{(1R)-2-[(6-{2-[(2,6-dichlorobenzyl)oxy]ethoxy}hexyl)amino]-1-hydroxyethyl}-2-(hydroxymethyl)phenoland related compounds disclosed in WO 03/024439, published on Mar. 27,2003; and pharmaceutically acceptable salts thereof. When employed, theβ₂-adrenoreceptor agonist will be present in the pharmaceuticalcomposition in a therapeutically effective amount. Typically, theβ₂-adrenoreceptor agonist will be present in an amount sufficient toprovide from about 0.05 μg to about 500 μg per dose.

Representative steroidal anti-inflammatory agents that can be used incombination with the compounds of this invention include, but are notlimited to, methyl prednisolone, prednisolone, dexamethasone,fluticasone propionate,6,9-difluoro-17-[(2-furanylcarbonyl)oxy]-11-hydroxy-16-methyl-3-oxoandrosta-1,4-diene-17-carbothioicacid S-fluoromethyl ester,6,9-difluoro-11-hydroxy-16-methyl-3-oxo-17-propionyloxy-androsta-1,4-diene-17-carbothioicacid S-(2-oxo-tetrahydrofuran-3S-yl)ester, beclomethasone esters (e.g.the 17-propionate ester or the 17,21-dipropionate ester), budesonide,flunisolide, mometasone esters (e.g. the furoate ester), triamcinoloneacetonide, rofleponide, ciclesonide, butixocort propionate, RPR-106541,ST-126 and the like, or pharmaceutically-acceptable salts thereof. Whenemployed, the steroidal anti-inflammatory agent will be present in thepharmaceutical composition in a therapeutically effective amount.Typically, the steroidal anti-inflammatory agent will be present in anamount sufficient to provide from about 0.05 μg to about 500 μg perdose.

Other suitable combinations include, for example, otheranti-inflammatory agents, e.g., NSAIDs (such as sodium cromoglycate;nedocromil sodium; phosphodiesterase (PDE) inhibitors (e.g.theophylline, PDE4 inhibitors or mixed PDE3/PDE4 inhibitors);leukotriene antagonists (e.g. monteleukast); inhibitors of leukotrienesynthesis; iNOS inhibitors; protease inhibitors, such as tryptase andelastase inhibitors; beta-2 integrin antagonists and adenosine receptoragonists or antagonists (e.g. adenosine 2a agonists); cytokineantagonists (e.g. chemokine antagonists such as, an interleukin antibody(IL antibody), specifically, an IL-4 therapy, an IL-13 therapy, or acombination thereof); or inhibitors of cytokine synthesis.

For example, representative phosphodiesterase-4 (PDE4) inhibitors ormixed PDE3/PDE4 inhibitors that can be used in combination with thecompounds of this invention include, but are not limited to cis4-cyano-4-(3-cyclopentyloxy-4-methoxyphenyl)cyclohexan-1-carboxylicacid,2-carbomethoxy-4-cyano-4-(3-cyclopropylmethoxy-4-difluoromethoxyphenyl)cyclohexan-1-one;cis-[4-cyano-4-(3-cyclopropylmethoxy-4-difluoromethoxyphenyl)cyclohexan-1-ol];cis-4-cyano-4-[3-(cyclopentyloxy)-4-methoxyphenyl]cyclohexane-1-carboxylicacid and the like, or pharmaceutically acceptable salts thereof. Otherrepresentative PDE4 or mixed PDE4/PDE3 inhibitors include AWD-12-281(elbion); NCS-613 (INSERM); D-4418 (Chiroscience and Schering-Plough);CI-1018 or PD-168787 (Pfizer); benzodioxole compounds disclosed inWO99/16766 (Kyowa Hakko); K-34 (Kyowa Hakko); V-11294A (Napp);roflumilast (Byk-Gulden); pthalazinone compounds disclosed in WO99/47505(Byk-Gulden); Pumafentrine (Byk-Gulden, now Altana); arofylline(Almirall-Prodesfarma); VM554/UM565 (Vernalis); T-440 (Tanabe Seiyaku);and T2585 (Tanabe Seiyaku).

Representative muscarinic antagonists (i.e., anticholinergic agents)that can be used in combination with, and in addition to, the compoundsof this invention include, but are not limited to, atropine, atropinesulfate, atropine oxide, methylatropine nitrate, homatropinehydrobromide, hyoscyamine (d, l) hydrobromide, scopolamine hydrobromide,ipratropium bromide, oxitropium bromide, tiotropium bromide,methantheline, propantheline bromide, anisotropine methyl bromide,clidinium bromide, copyrrolate (Robinul), isopropamide iodide,mepenzolate bromide, tridihexethyl chloride (Pathilone), hexocycliummethylsulfate, cyclopentolate hydrochloride, tropicamide,trihexyphenidyl hydrochloride, pirenzepine, telenzepine, AF-DX 116 andmethoctramine and the like, or a pharmaceutically acceptable saltthereof; or, for those compounds listed as a salt, alternatepharmaceutically acceptable salt thereof.

Representative antihistamines (i.e., H₁-receptor antagonists) that canbe used in combination with the compounds of this invention include, butare not limited to, ethanolamines, such as carbinoxamine maleate,clemastine fumarate, diphenylhydramine hydrochloride and dimenhydrinate;ethylenediamines, such as pyrilamine amleate, tripelennaminehydrochloride and tripelennamine citrate; alkylamines, such aschlorpheniramine and acrivastine; piperazines, such as hydroxyzinehydrochloride, hydroxyzine pamoate, cyclizine hydrochloride, cyclizinelactate, meclizine hydrochloride and cetirizine hydrochloride;piperidines, such as astemizole, levocabastine hydrochloride, loratadineor its descarboethoxy analogue, terfenadine and fexofenadinehydrochloride; azelastine hydrochloride; and the like, or apharmaceutically acceptable salt thereof; or, for those compounds listedas a salt, alternate pharmaceutically acceptable salt thereof.

Suitable doses for the other therapeutic agents administered incombination with a compound of the invention are in the range of about0.05 g/day to about 100 mg/day.

The following formulations illustrate representative pharmaceuticalcompositions of the present invention:

FORMULATION EXAMPLE A

A dry powder for administration by inhalation is prepared as follows:Ingredients Amount Compound of the invention 0.2 mg Lactose  25 mg

-   -   Representative Procedure: The compound of the invention is        micronized and then blended with lactose. This blended mixture        is then loaded into a gelatin inhalation cartridge. The contents        of the cartridge are administered using a powder inhaler.

FORMULATION EXAMPLE B

A dry powder formulation for use in a dry powder inhalation device isprepared as follows:

Representative Procedure: A pharmaceutical composition is preparedhaving a bulk formulation ratio of micronized compound of the inventionto lactose of 1:200. The composition is packed into a dry powderinhalation device capable of delivering between about 10 μg and about100 μg of the compound of the invention per dose.

FORMULATION EXAMPLE C

A dry powder for administration by inhalation in a metered dose inhaleris prepared as follows:

Representative Procedure: A suspension containing 5 wt. % of a compoundof the invention and 0.1 wt. % lecithin is prepared by dispersing 10 gof the compound of the invention as micronized particles with mean sizeless than 10 μm in a solution formed from 0.2 g of lecithin dissolved in200 mL of demineralized water. The suspension is spray dried and theresulting material is micronized to particles having a mean diameterless than 1.5 μm. The particles are loaded into cartridges withpressurized 1,1,1,2-tetrafluoroethane.

FORMULATION EXAMPLE D

A pharmaceutical composition for use in a metered dose inhaler isprepared as follows:

Representative Procedure: A suspension containing 5% compound of theinvention, 0.5% lecithin, and 0.5% trehalose is prepared by dispersing 5g of active ingredient as micronized particles with mean size less than10 m in a colloidal solution formed from 0.5 g of trehalose and 0.5 g oflecithin dissolved in 100 mL of demineralized water. The suspension isspray dried and the resulting material is micronized to particles havinga mean diameter less than 1.5 μm. The particles are loaded intocanisters with pressurized 1,1,1,2-tetrafluoroethane.

FORMULATION EXAMPLE E

A pharmaceutical composition for use in a nebulizer inhaler is preparedas follows:

Representative Procedure: An aqueous aerosol formulation for use in anebulizer is prepared by dissolving 0.1 mg of the compound of theinvention in 1 mL of a 0.9% sodium chloride solution acidified withcitric acid. The mixture is stirred and sonicated until the activeingredient is dissolved. The pH of the solution is adjusted to a valuein the range of from 3 to 8 by the slow addition of NaOH.

FORMULATION EXAMPLE F

Hard gelatin capsules for oral administration are prepared as follows:Ingredients Amount Compound of the invention 250 mg Lactose(spray-dried) 200 mg Magnesium stearate  10 mg

-   -   Representative Procedure: The ingredients are thoroughly blended        and then loaded into a hard gelatin capsule (460 mg of        composition per capsule).

FORMULATION EXAMPLE G

A suspension for oral administration is prepared as follows: IngredientsAmount Compound of the invention 1.0 g Fumaric acid 0.5 g Sodiumchloride 2.0 g Methyl paraben 0.15 g Propyl paraben 0.05 g Granulatedsugar 25.5 g Sorbitol (70% solution) 12.85 g Veegum k (Vanderbilt Co.)1.0 g Flavoring 0.035 mL Colorings 0.5 mg Distilled water q.s. to 100 mL

-   -   Representative Procedure: The ingredients are mixed to form a        suspension containing 100 mg of active ingredient per 10 mL of        suspension.

FORMULATION EXAMPLE H

An injectable formulation is prepared as follows: Ingredients AmountCompound of the invention 0.2 g Sodium acetate buffer solution (0.4 M)2.0 mL HCl (0.5 N) or NaOH (0.5 N) q.s. to pH 4 Water (distilled,sterile) q.s. to 20 mL

-   -   Representative Procedure: The above ingredients are blended and        the pH is adjusted to 4±0.5 using 0.5 N HCl or 0.5 N NaOH.

Utility

The biphenyl derivatives of this invention possess both β₂ adrenergicreceptor agonist and muscarinic receptor antagonist activity andtherefore, such compounds are useful for treating medical conditionsmediated by β₂ adrenergic receptors or muscarinic receptors, i.e.,medical conditions that are ameliorated by treatment with a β₂adrenergic receptor agonist or a muscarinic receptor antagonist. Suchmedical conditions include, by way of example, pulmonary disorders ordiseases associated with reversible airway obstruction, such as chronicobstructive pulmonary disease (e.g., chronic and wheezy bronchitis andemphysema), asthma, pulmonary fibrosis and the like. Other conditionswhich may be treated include premature labor, depression, congestiveheart failure, skin diseases (e.g., inflammatory, allergic, psoriaticand proliferative skin diseases, conditions where lowering pepticacidity is desirable (e.g., peptic and gastric ulceration) and musclewasting disease.

Accordingly, in one embodiment, this invention is directed to a methodfor treating a pulmonary disorder, the method comprising administeringto a patient in need of treatment a therapeutically effective amount ofa compound of formula I or a pharmaceutically acceptable salt or solvateor stereoisomer thereof. When used to treat a pulmonary disorder, thecompounds of this invention will typically be administered by inhalationin multiple doses per day, in a single daily dose or a single weeklydose. Generally, the dose for treating a pulmonary disorder will rangefrom about 10 μg/day to about 200 μg/day.

When administered by inhalation, the compounds of this inventiontypically have the effect of providing bronchodilation. Accordingly, inanother of its method aspects, this invention is directed to a method ofproviding bronchodilation in a patient, the method comprisingadministering to a patient requiring bronchodilation a therapeuticallyeffective amount of a compound of formula I or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof. Generally, the dosefor providing bronchodilation will range from about 10 μg/day to about200 μg/day.

In one embodiment, this invention is directed to a method of treatingchronic obstructive pulmonary disease or asthma, the method comprisingadministering to a patient in need of treatment a therapeuticallyeffective amount of a compound of formula I or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof. When used to treat aCOPD or asthma, the compounds of this invention will typically beadministered by inhalation in multiple doses per day or in a singledaily dose. Generally, the dose for treating COPD or asthma will rangefrom about 10 μg/day to about 200 μg/day.

As used herein, COPD includes chronic obstructive bronchitis andemphysema (see, for example, Barnes, Chronic Obstructive PulmonaryDisease, N Engl J Med 2000: 343:269-78).

When used to treat a pulmonary disorder, the compounds of this inventionare optionally administered in combination with other therapeuticagents. In particular, by combining the compounds of this invention witha steroidal anti-inflammatory agent (e.g. a corticosteroid), thepharmaceutical compositions of this invention can provide tripletherapy, i.e., β₂ adrenergic receptor agonist, muscarinic receptorantagonist and anti-inflammatory activity, using only two activecomponents. Since pharmaceutical compositions containing two activecomponents are typically easier to formulate compared to compositionscontaining three active components, such two component compositionsprovide a significant advantage over compositions containing threeactive components. Accordingly, in a particular embodiment, thepharmaceutical compositions and methods of this invention furthercomprise a therapeutically effective amount of a steroidalanti-inflammatory agent.

Compounds of this invention exhibit both muscarinic receptor antagonistand β₂ adrenergic receptor agonist activity. Accordingly, among otherproperties, compounds of particular interest are those that demonstratean inhibitory constant K_(i) value for binding at the M₃ muscarinicreceptor and an EC₅₀ value for β₂ adrenergic receptor agonist activityof less than about 100 nM; particularly less than 10 nM. Among thesecompounds, compounds of special interest include those having muscarinicactivity, expressed in terms of the inhibitory constant K_(i) forbinding at the M₃ muscarinic receptor, that is about equal to thecompound's β₂ adrenergic agonist activity, expressed in terms of thehalf maximal effective concentration EC₅₀, as determined in the in vitroassays described herein, or in similar assays. For example, compounds ofparticular interest are those having a ratio of the inhibitory constantK_(i) for the M₃ muscarinic receptor to the EC₅₀ for the β₂ adrenergicreceptor ranging from about 30:1 to about 1:30; including about 20:1 toabout 1:20; such as about 10:1 to about 1:10.

In one of its method aspects, the present invention also provides amethod for treating a pulmonary disorder, the method comprisingadministering to a patient in need of treatment a therapeuticallyeffective amount of a compound having both muscarinic receptorantagonist and β₂ adrenergic receptor agonist activity. In a particularembodiment of this method, the compound administered has an inhibitoryconstant K_(i) for the M₃ muscarinic receptor that is less than about100 nM and a half maximal effective concentration EC₅₀ for agonism atthe β₂ adrenergic receptor that is less than about 100 nM. In anotherembodiment, the method for treating a pulmonary disorder comprisesadministering a therapeutically effective amount of a compound for whichthe ratio of the inhibitory constant K_(i) for the M₃ muscarinicreceptor to the EC₅₀ for agonism of the β₂ adrenergic receptor isbetween about 30:1 and about 1:30.

Since compounds of this invention possess both β₂ adrenergic agonistactivity and muscarinic receptor antagonist activity, such compounds arealso useful as research tools for investigating or studying biologicalsystems or samples having β₂ adrenergic receptors or muscarinicreceptors, or for discovering new compounds having both β₂ adrenergicagonist activity and muscarinic receptor antagonist activity. Suchbiological systems or samples may comprise β₂ adrenergic receptorsand/or muscarinic receptors. Any suitable biological system or samplehaving β₂ adrenergic and/or muscarinic receptors may be employed in suchstudies which may be conducted either in vitro or in vivo.Representative biological systems or samples suitable for such studiesinclude, but are not limited to, cells, cellular extracts, plasmamembranes, tissue samples, mammals (such as mice, rats, guinea pigs,rabbits, dogs, pigs, etc.), and the like.

In this embodiment, a biological system or sample comprising a β₂adrenergic receptor or a muscarinic receptor is contacted with a β₂adrenergic receptor-agonizing or muscarinic receptor-antagonizing amountof a compound of this invention. The effects are then determined usingconventional procedures and equipment, such as radioligand bindingassays and functional assays. Such functional assays includeligand-mediated changes in intracellular cyclic adenosine monophosphate(cAMP), ligand-mediated changes in activity of the enzyme adenylylcyclase (which synthesizes cAMP), ligand-mediated changes inincorporation of guanosine 5′-O—(-thio)triphosphate ([³⁵S]GTP S) intoisolated membranes via receptor catalyzed exchange of [³⁵S]GTP S forGDP, ligand-mediated changes in free intracellular calcium ions(measured, for example, with a fluorescence-linked imaging plate readeror FLIPR® from Molecular Devices, Inc.). A compound of this inventionwill agonize or cause activation of a β₂ adrenergic receptor andantagonize or decrease the activation of muscarinic receptors in any ofthe functional assays listed above, or assays of a similar nature. Theamount of compound used in these studies will typically range from about0.1 nanomolar to about 100 nanomolar.

Additionally, the compounds of this invention can be used as researchtools for discovering new compounds that have both a β₂ adrenergicreceptor agonist and muscarinic receptor antagonist activity. In thisembodiment, a β₂ adrenergic receptor and muscarinic receptor bindingdata (for example, as determined by in vitro radioligand displacementassays) for a test compound or a group of test compounds is compared tothe β₂ adrenergic receptor and muscarinic receptor binding data for acompound of this invention to identify those test compounds that haveabout equal or superior β₂ adrenergic receptor and/or muscarinicreceptor binding, if any. This aspect of the invention includes, asseparate embodiments, both the generation of comparison data (using theappropriate assays) and the analysis of the test data to identify testcompounds of interest.

In some cases, compounds of this invention may possess either weakmuscarinic receptor antagonist activity or weak β₂ adrenergic receptoragonist activity. In these cases, those of ordinary skill in the artwill recognize that such compounds still have utility as primarilyeither a β₂ adrenergic receptor agonist or a muscarinic receptorantagonist, respectively.

The properties and utility of the compounds of this invention can bedemonstrated using various in vitro and in vivo assays well-known tothose skilled in the art. For example, representative assays aredescribed in further detail in the following Examples.

EXAMPLES

The following Preparations and Examples are provided to illustratespecific embodiments of this invention. These specific embodiments,however, are not intended to limit the scope of this invention in anyway unless specifically indicated.

The following abbreviations have the following meanings unless otherwiseindicated and any other abbreviations used herein and not defined havetheir standard meaning:

-   -   AC adenylyl cyclase    -   Ach acetylcholine    -   ATCC American Type Culture Collection    -   BSA bovine serum albumin    -   cAMP 3′-5′ cyclic adenosine monophosphate    -   CHO Chinese hamster ovary    -   cM₅ cloned chimpanzee M₅ receptor    -   DCM dichloromethane (i.e., methylene chloride)    -   DIPEA N,N-diisopropylethylamine    -   dPBS Dulbecco's phosphate buffered saline    -   DMEM Dulbecco's Modified Eagle's Medium    -   DMSO dimethyl sulfoxide    -   EDTA ethylenediaminetetraacetic acid    -   Emax maximal efficacy    -   EtOAc ethyl acetate    -   EtOH ethanol    -   FBS fetal bovine serum    -   FLIPR fluorometric imaging plate reader    -   Gly glycine    -   HATU O-(7-azabenzotriazol-1-yl-N,N,N′,N′-tetramethyluronium        hexafluorophosphate    -   HBSS Hank's buffered salt solution    -   HEK human embryonic kidney cells    -   HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid    -   hM₁ cloned human M₁ receptor    -   hM₂ cloned human M₂ receptor    -   hM₃ cloned human M₃ receptor    -   hM₄ cloned human M₄ receptor    -   hM₅ cloned human M₅ receptor    -   HPLC high-performance liquid chromatography    -   IBMX 3-isobutyl-1-methylxanthine    -   % Eff % efficacy    -   PBS phosphate buffered saline    -   PyBOP benzotriazol-1-yloxytripyrrolidinophosphonium        hexafluorophosphate    -   rpm rotations per minute    -   TFA trifluoroacetic acid    -   THF tetrahydrofuran    -   Tris tris(hydroxymethyl)aminomethane

Unless noted otherwise, reagents, starting materials and solvents werepurchased from commercial suppliers (such as Aldrich, Fluka, Sigma andthe like) and were used without further purification.

In the examples described below, HPLC analysis was conducted using anAgilent (Palo Alto, Calif.) Series 1100 instrument with Zorbax Bonus RP2.1×50 mm columns, supplied by Agilent, (a C14 column), having a 3.5micron particle size. Detection was by UV absorbance at 214 nm. HPLC10-70 data was obtained with a flow rate of 0.5 mL/minute of 10%-70% Bover 6 minutes. Mobile phase A was 2%-98%-0.1% ACN—H₂O-TFA; and mobilephase B was 90%-10%-0.1% ACN—H₂O-TFA. Using the mobile phases A and Bdescribed above, HPLC 5-35 data and HPLC 10-90 data were obtained with a5 minute gradient.

Liquid chromatography mass spectrometry (LCMS) data were obtained withan Applied Biosystems (Foster City, Calif.) model API-150EX instrument.LCMS 10-90 data was obtained with a 10%-90% mobile phase B over a 5minute gradient.

Small scale purification was conducted using an API 150EX PrepWorkstation system from Applied Biosystems. The mobile phase was A:water+0.05% v/v TFA; and B: acetonitrile+0.05% v/v TFA. For arrays(typically about 3 to 50 mg recovered sample size) the followingconditions were used: 20 mL/min flow rate; 15 min gradients and a 20mm×50 mm Prism RP column with 5 micron particles (ThermoHypersil-Keystone, Bellefonte, Pa.). For larger scale purifications(typically greater than 100 mg crude sample), the following conditionswere used: 60 mL/min flow rate; 30 min gradients and a 41.4 mm×250 mmMicrosorb BDS column with 10 micron particles (Varian, Palo Alto,Calif.).

The specific rotation for chiral compounds (indicated as [α]²⁰ _(D)) wasmeasured using a Jasco Polarimeter (Model P-1010) with a tungstenhalogen light source and a 589 nm filter at 20° C. Samples of testcompounds were typically measured at 1 mg/mL water.

Preparation 1

N-1,1′-Biphenyl-2-yl-N′-4-(1-benzyl)piperidinylurea

Biphenyl-2-isocyanate (50 g, 256 mmol) was dissolved in acetonitrile(400 mL) at ambient temperature. After cooling to 0° C., a solution of4-amino-N-benzylpiperidine (48.8 g, 256 mmol) in acetonitrile (400 mL)was added over 5 min. A precipitate was observed immediately. After 15min, acetonitrile (600 mL) was added, and the resultant viscous mixturewas stirred for 12 h at 35° C. The solids were then filtered off andwashed with cold acetonitrile, then dried under vacuum, yielding thetitle compound (100 g, 98% yield). MS m/z: [M+H⁺] calcd for C₂₅H₂₇N₃O386.22. found 386.3.

Preparation 2

N-1,1′-Biphenyl-2-yl-N′-4-piperidinylurea

The product of Preparation 1 (20 g, 52 mmol) was dissolved in a mixtureof anhydrous methanol and anhydrous DMF (3:1, 800 mL). Aqueoushydrochloric acid (0.75 mL of 37% conc. solution, 7.6 mmol) was addedand nitrogen gas was bubbled through the solution vigorously for 20 min.Pearlman's catalyst (Pd(OH)₂, 5 g) was added under a stream of nitrogen,before placing the reaction mixture under a hydrogen atmosphere(balloon). The reaction mixture was allowed to stir for 4 days and wasthen passed twice through pads of Celite to remove the catalyst. Thesolvent was then removed under reduced pressure to yield the titlecompound (13 g, 85% yield). MS m/z: [M+H⁺] calcd for C₁₈H₂₁N₃O 296.17.found 296.0.

Alternatively, N-1,1′-biphenyl-2-yl-N′-4-piperidinylurea was synthesizedby heating together biphenyl-2-isocyanate (50 g, 256 mmol) and4-amino-N-benzylpiperidine (51.1 g, 269 mmol) at 70° C. for 12 h (thereaction was monitored by LCMS). The reaction mixture was cooled to 50°C. and ethanol (500 mL) added, followed by slow addition of 6Mhydrochloric acid (95 mL). The reaction mixture was cooled to roomtemperature. Ammonium formate (48.4 g, 768 mmol) was added to thereaction mixture and nitrogen gas bubbled through the solutionvigorously for 20 min, before adding palladium (10 wt. % (dry basis) onactivated carbon) (10 g). The reaction mixture was heated at 40° C. for12 h, and then filtered through a pad of Celite and the solvent wasremoved under reduced pressure. To the crude residue was added 1Mhydrochloric acid (20 mL) and 10N sodium hydroxide was added to adjustthe pH to 12. The aqueous layer was extracted with ethyl acetate (2×80mL), dried (magnesium sulfate) and solvent removed under reducedpressure to give the title compound as a solid (71.7 g, 95% yield). MSm/z: [M+H⁺] calcd for C₁₈H₂₁N₃O 296.17. found 296.0.

Preparation 3

N-1,1′-Biphenyl-2-yl-N′-4-[1-(9-hydroxynonyl)]piperidinylurea

9-Bromo-1-nonanol (4.84 g, 21.7 mmol) was added to a stirred solution ofthe product of Preparation 2 (5.8 g, 19.7 mmol) anddiisopropylethylamine (10.29 mL, 59.1 mmol) in acetonitrile (99 mL) at50° C. The reaction mixture was heated at 50° C. for 8 h. The reactionmixture was then allowed to cool and the solvent was removed underreduced pressure. The residue was dissolved in dichloromethane (100 mL),washed with saturated aqueous sodium bicarbonate (2×50 mL) and dried(magnesium sulfate). The solvent was removed under reduced pressure. Thecrude product was purified by flash chromatography(dichloromethane:methanol:ammonia system) to yield the title compound(7.1 g, 16.2 mmol, 82% yield).

Preparation 4

N-1,1′-Biphenyl-2-yl-N′-4-[1-(9-oxononyl)]piperidinylurea

Dimethyl sulfoxide (490 μL, 6.9 mmol), followed by diisopropylethylamine(324 μL, 3.45 mmol) was added to a solution of the product ofPreparation 3 (500 mg, 1.15 mmol) in dichloromethane (11.5 mL) at −10°C. under an atmosphere of nitrogen. The reaction mixture was stirred at−15° C. for 15 min, and then sulfur trioxide pyridine complex was addedportionwise (549 mg, 3.45 mmol). The reaction mixture was stirred at−15° C. for 1 h, and then water (10 mL) was added. The organic phase wasthen separated, washed with water (10 mL), and dried (sodium sulfate).The solvent was removed under reduced pressure to give the titlecompound (475 mg, 1.09 mmol, 95% yield). HPLC (10-70) R_(t)=3.39.

Preparation 5

N-1,1′-Biphenyl-2-yl-N′-4-[1-(9-aminononyl)]piperidinylurea

Palladium (10 wt. % (dry basis) on activated carbon) (1.5 g) was addedto a stirred solution of the product of Preparation 4 (1.58 g, 3.63mmol) and benzylamine (516 μL, 4.72 mmol) in methanol (36.3 mL). Thereaction mixture was placed under an atmosphere of hydrogen. Afterstirring for 12 h, the reaction mixture was filtered through a pad ofCelite and washed with methanol (10 mL). The solvent was removed underreduced pressure to give the title compound (1.50 g, 3.45 mmol, 95%yield). HPLC (10-70) R_(t)=2.35; MS m/z: [M+H⁺] calcd for C₂₇H₄₀N₄O₁437.06. found 437.5.

Preparation 6

8-Benzyloxy-5-(2,2-dihydroxyacetyl)-1H-quinolin-2-one (a)8-Acetoxy-1H-quinolin-2-one

8-Hydroxyquinoline-N-oxide (160.0 g, 1.0 mol), commercially-availablefrom Aldrich, Milwaukee, Wis., and acetic anhydride (800 mL, 8.4 mol)were heated at 100° C. for 3 h and then cooled in ice. The product wascollected on a Buchner funnel, washed with acetic anhydride (2×100 mL)and dried under reduced pressure to give 8-acetoxy-1H-quinolin-2-one(144 g) as a solid.

(b) 5-Acetyl-8-hydroxy-1H-quinolin-2-one

A slurry of aluminum chloride (85.7 g, 640 mmol) in 1,2-dichloroethane(280 mL) was cooled in ice, and the product of step (a) (56.8 g, 280mmol) was added. The mixture was warmed to room temperature and thenheated at 85° C. After 30 min, acetyl chloride (1.5 mL, 21 mmol) wasadded and the mixture was heated an additional 60 min. The reactionmixture was then cooled and added to 1N hydrochloric acid (3 L) at 0° C.with good stirring. After stirring for 2 h, the solids were collected ona Buchner funnel, washed with water (3×250 mL) and dried under reducedpressure. The crude product isolated from several batches (135 g) wascombined and triturated with dichloromethane (4 L) for 6 h. The productwas collected on a Buchner funnel and dried under reduced pressure togive 5-acetyl-8-hydroxy-2(1H)-quinolinone (121 g).

(c) 5-Acetyl-8-benzyloxy-1H-quinolin-2-one

To the product of step (b) (37.7 g, 186 mmol) was addedN,N-dimethylformamide (200 mL) and potassium carbonate (34.5 g, 250mmol) followed by benzyl bromide (31.8 g, 186 mmol). The mixture wasstirred at room temperature for 2.25 hour and then poured into saturatedsodium chloride (3.5 L) at 0° C. and stirred for 1 hour. The product wascollected and dried on a Buchner funnel for 1 hour, and the resultingsolids were dissolved in dichloromethane (2 L) and this mixture wasdried over sodium sulfate. The solution was filtered through a pad ofCelite which was then washed with dichloromethane (5×200 mL). Thecombined filtrate was then concentrated to dryness and the resultingsolids were triturated with ether (500 mL) for 2 h. The product wascollected on a Buchner funnel, washed with ether (2×250 mL) and driedunder reduced pressure to give 5-acetyl-8-benzyloxy-1H-quinolin-2-one(44 g) as a powder.

(d) 8-Benzyloxy-5-(2,2-dihydroxyacetyl)-1H-quinolin-2-one

To a slurry of the product of step (c) (10.0 g, 34.1 mmol) in DMSO (60mL) was added a 48% w/w hydrobromic acid solution (11.8 mL, 102.3 mmol).The mixture was warmed to 60° C. for 16 h then allowed to cool to roomtemperature. Water (100 mL) was added and the resulting slurry stirredat room temperature for 0.5 h before being cooled to 0° C. The productwas collected on a Buchner funnel then dried under reduced pressure togive 8-benzyloxy-5-(2,2-dihydroxyacetyl)-1H-quinolin-2-one (12.2 g) as asolid.

Preparation 7

1-(1-{9-[2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]nonyl}piperidin-4-yl)-3-biphenyl-2-ylurea

The products of Preparation 5 (183 mg, 0.42 mmol) and Preparation 6 (149mg, 0.46 mmol) were stirred in dichloroethane (4.2 mL) at ambienttemperature for 2 h. Sodium triacetoxyborohydride (267 mg, 1.26 mmol)was then added and the reaction mixture was stirred for a further 12 h.The reaction mixture was then diluted with dichloromethane (10 mL),washed with saturated aqueous sodium bicarbonate (10 mL), dried(magnesium sulfate) and the solvent was removed under reduced pressure.The crude reaction mixture was purified by flash chromatography (5-10%methanol in dichloromethane, 0.5% ammonium hydroxide) to give the titlecompound (144 mg, 0.20 mmol, 48% yield). HPLC (10-70) R_(t)=3.48; MSm/z: [M+H⁺] calcd for C₄₅H₅₅N₅O₄ 730.4. found 730.7.

Example 11-Biphenyl-2-yl-3-(1-{9-[2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-yl)urea

Palladium (10 wt. % (dry basis) on activated carbon) (63 mg) was addedto a stirred solution of the product of Preparation 7 (144 mg, 0.20mmol) in methanol (2 mL) and the reaction mixture was placed under anatmosphere of hydrogen. After 12 h stirring, the reaction mixture wasfiltered through a pad of Celite, washed with methanol (2 mL) and thenthe solvent was removed under reduced pressure. The resulting residuewas purified by preparative HPLC to give the title compound (10 mg).HPLC (10-70) R_(t)=2.8; MS m/z: [M+H⁺] calcd for C₃₈H₄₉N₅O₄ 640.3. found640.5.

Preparation 8

Biphenyl-2-ylcarbamic Acid Piperidin-4-yl Ester

Biphenyl-2-isocyanate (97.5 g, 521 mmol) and4-hydroxy-1-benzylpiperidine (105 g, 549 mmol), bothcommercially-available from Aldrich, Milwaukee, Wis., were heatedtogether at 70° C. for 12 h, during which time the formation ofbiphenyl-2-ylcarbamic acid 1-benzylpiperidin-4-yl ester was monitored byLCMS. The reaction mixture was then cooled to 50° C. and ethanol (1 L)was added, and then 6M hydrochloric acid (191 mL) was added slowly. Thereaction mixture was then cooled to ambient temperature and ammoniumformate (98.5 g, 1.56 mol) was added and nitrogen gas was bubbledthrough the solution vigorously for 20 min. Palladium (10 wt. % (drybasis) on activated carbon) (20 g) was then added. The reaction mixturewas heated at 40° C. for 12 h and then filtered through a pad of Celite.The solvent was then removed under reduced pressure and 1M hydrochloricacid (40 mL) was added to the crude residue. Sodium hydroxide (10N) wasthen added to adjust the pH to 12. The aqueous layer was extracted withethyl acetate (2×150 mL) and dried (magnesium sulfate), and then thesolvent was removed under reduced pressure to give the title compound(155 g, 100%). HPLC (10-70) R_(t)=2.52; MS m/z: [M+H⁺] calc'd forC₁₈H₂₀N₂O₂ 297.15. found 297.3.

Preparation 9

N,N-(Di-tert-butoxycarbonyl)-9-bromononylamine

A solution of di-tert-butoxycarbonylamine (3.15 g, 14.5 mmol) inN,N-dimethylformamide (0.28 mL) was cooled to 0° C. for about 10 min.Sodium hydride, 60% in mineral oil (0.58 g, 14.5 mmol) was added and thereaction mixture was stirred at 0° C. for 10 min. The reaction mixturewas removed from the ice bath and allowed to warm to room temperaturefor about 30 min. The reaction mixture was then cooled back down to 0°C. and a solution of 1,9-dibromononane (2.46 mL, 12.1 mmol) indimethylformamide (100 mL) was added. The reaction mixture was stirredovernight at room temperature. After 24 h, MS analysis showed that thereaction was completed. The reaction mixture was concentrated to drynessand diluted with ethyl acetate (100 mL). The organic layer was washedwith saturated sodium bicarbonate (2×100 mL), brine (100 mL), dried(magnesium sulfate) and concentrated under reduced pressure to yield thecrude product, which was purified by chromatography on silica gel using5% ethyl acetate in hexanes to afford the title compound. MS m/z: [M+H⁺]calcd for C₁₉H₃₆N₁O₄Br 423.18. found 423.

Preparation 10

Biphenyl-2-ylcarbamic Acid1-(9-Di-tert-butoxycarbonylamino)nonyl]piperidin-4-yl Ester

A mixture of 1:1 acetonitrile and N,N-dimethylformamide (50 mL) wasadded to the products of Preparation 8 (3.0 g, 10.1 mmol) andPreparation 9 (5.1 g, 12.2 mmol) and triethylamine (1.42 mL, 10.1 mmol).The reaction mixture was stirred at ambient temperature for 24 h and wasmonitored by LCMS analysis. The reaction mixture was then concentratedand diluted with ethyl acetate (50 mL). The organic layer was washedwith saturated sodium bicarbonate (2×50 mL) and brine (50 mL). Theorganic phase was then dried over magnesium sulfate and concentrated toyield 6.5 g of crude oil. The oil was purified by chromatography onsilica gel using 1:1 hexanes/ethyl acetate to provide the title compound(3 g). MS m/z: [M+H⁺] calcd for C₃₇H₅₅N₃O₆ 638.41. found 639.

Preparation 11

Biphenyl-2-ylcarbamic Acid 1-(9-Aminononyl)piperidin-4-yl Ester

Trifluoroacetic acid (11 mL) was added to a solution of the product ofPreparation 10 (7.2 g, 11.3 mmol) in dichloromethane (56 mL). After 2 h,LCMS analysis showed that the reaction was completed. The reactionmixture was then concentrated to dryness and diluted with ethyl acetate(75 mL). Sodium hydroxide (1N) was then added until the pH of themixture reached 14. The organic phase was then collected and washed withsaturated sodium bicarbonate (2×50 mL) and brine (50 mL). The organicphase was then dried over magnesium sulfate and concentrated to providethe title compound (5.5 g). MS m/z: [M+H⁺] calcd for C₂₇H₃₉N₃O₂ 438.30.found 439.

Preparation 12

Biphenyl-2-ylcarbamic Acid1-{9-[2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]nonyl}piperidin-4-ylEster

The product of Preparation 11 (196 mg, 0.43 mmol) was dissolved indichloroethane (4 mL) and sodium triacetoxyborohydride (101 mg, 0.48mmol) was added. The reaction mixture was stirred at ambient temperaturefor about 10 min. and then8-benzyloxy-5-(2,2-dihydroxyacetyl)-1H-quinolin-2-one (Preparation 6)(141 mg, 0.43 mmol) was added. LCMS analysis showed that the reactionwas completed after 2 h. Methanol (1 mL) was added to the reactionmixture and then sodium borohydride (18 mg, 0.48 mmol) was added slowly.After 1 hour, LCMS analysis showed that the reaction was completed. Thereaction mixture was then quenched with aqueous ammonium chloride andthis mixture was extracted with dichloromethane. The organic phase waswashed with saturated sodium bicarbonate (2×50 mL) and brine (10 mL).The organic phase was then dried over magnesium sulfate and concentratedto provide 315 mg of a yellow solid. The solid was purified by silicagel chromatography using 10% methanol in dichloromethane to afford thetitle compound (64 mg). MS m/z: [M+H⁺] calcd for C₄₃H₅₅N₄O₅ 730.40.found 731.

Example 2 Biphenyl-2-ylcarbamic Acid1-{9-[2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

A solution of the product of Preparation 12 (64 mg, 0.09 mmol) inmethanol (450 mL) was flushed with nitrogen. Palladium on carbon (10%,10 mg) was then added and the reaction mixture was placed under aballoon containing hydrogen and stirred. LCMS analysis showed that thereaction was completed after 9 h. The reaction mixture was then filteredand the filtrate was concentrated to provide a yellow crispy solid. Thesolid was purified by preparative HPLC (5-35 over 60 min) to afford thetitle compound (19 mg). MS m/z: [M+H⁺] calcd for C₃₈H₄₈N₄O₅ 641.36.found 641.

Preparation 13

8-Benzyloxy-5-[(R)-2-bromo-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one(a) 8-Benzyloxy-5-(2-Bromoacetyl)-1H-quinolin-2-one

5-Acetyl-8-benzyloxy-1H-quinolin-2-one (Preparation 6) (20.0 g, 68.2mmol) was dissolved in dichloromethane (200 mL) and cooled to 0° C.Boron trifluoride diethyl etherate (10.4 mL, 82.0 mmol) was added viasyringe and the mixture was warmed to room temperature to give a thicksuspension. The suspension was heated at 45° C. (oil bath) and asolution of bromine (11.5 g, 72.0 mmol) in dichloromethane (100 mL) wasadded over 40 min. The mixture was kept at 45° C. for an additional 15min and then cooled to room temperature. The mixture was concentratedunder reduced pressure and then triturated with 10% aqueous sodiumcarbonate (200 mL) for 1 hour. The solids were collected on a Buchnerfunnel, washed with water (4×100 mL) and dried under reduced pressure.The product of two runs was combined for purification. The crude product(52 g) was triturated with 50% methanol in chloroform (500 mL) for 1hour. The product was collected on a Buchner funnel and washed with 50%methanol in chloroform (2×50 mL) and methanol (2×50 mL). The solid wasdried under reduced pressure to give the title compound (34.1 g) as apowder.

(b) 8-Benzyloxy-5-((R)-2-bromo-1-hydroxyethyl)-1H-quinolin-2-one

(R)-(+)-α,α-Diphenylprolinol (30.0 g, 117 mmol) and trimethylboroxine(11.1 mL, 78 mmol) were combined in toluene (300 mL) and stirred at roomtemperature for 30 min. The mixture was placed in a 150° C. oil bath andliquid was distilled off. Toluene was added in 20 mL aliquots anddistillation was continued for 4 h. A total of 300 mL toluene was added.The mixture was then cooled to room temperature. A 500 μL aliquot wasevaporated to dryness and weighed (246 mg) to determine that theconcentration of catalyst was 1.8 M.

8-Benzyloxy 5-(2-bromoacetyl)-1H-quinolin-2-one (90.0 g, 243 mmol) wasplaced under nitrogen and tetrahydrofuran (900 mL) was added followed bythe catalyst described above (1.8 M in toluene, 15 mL, 27 mmol). Thesuspension was cooled to −10±5° C. in an ice/isopropanol bath. Borane(1.0 M in THF, 294 mL, 294 mmol) was added over 4 h. The reaction wasthen stirred an additional 45 min at −10° C. and then methanol (250 mL)was added slowly. The mixture was concentrated under vacuum and theresidue was dissolved in boiling acetonitrile (1.3 L), filtered whilehot and then cooled to room temperature. The crystals were filtered,washed with acetonitrile and dried under vacuum to give the titlecompound (72.5 g, 196 mmol, 81% yield, 95% ee, 95% pure by HPLC).

(c)8-Benzyloxy-5-[(R)-2-bromo-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one

To the product of step (b) (70.2 g, 189 mmol) was addedN,N-dimethylformamide (260 mL) and this mixture was cooled in an icebath under nitrogen. 2,6-Lutidine (40.3 g, 376 mmol) was added over 5min and then tert-butyldimethylsilyl trifluoromethanesulfonate (99.8 g,378 mmol) was added slowly while maintaining the temperature below 20°C. The mixture was allowed to warm to room temperature for 45 min.Methanol (45 mL) was added to the mixture dropwise over 10 min and themixture was partitioned between ethyl acetate/cyclohexane (1:1, 500 mL)and water/brine (1:1, 500 mL). The organics were washed twice more withwater/brine (1:1, 500 mL each). The combined organics were evaporatedunder reduced pressure to give a light yellow oil. Two separate portionsof cyclohexane (400 mL) were added to the oil and distillation continueduntil a thick white slurry was formed. Cyclohexane (300 mL) was added tothe slurry and the resulting white crystals were filtered, washed withcyclohexane (300 mL) and dried under reduced pressure to give the titlecompound (75.4 g, 151 mmol, 80% yield, 98.6% ee).

Preparation 14

Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]nonyl}piperidin-4-ylEster

The product of Preparation 13 (3.9 g, 8.17 mmol) was added to a solutionof the product of Preparation 11 (5.0 g, 11.4 mmol) in THF (20 mL),followed by sodium bicarbonate (2.0 g, 24.5 mmol) and sodium iodide (1.8g, 12.2 mmol). The reaction mixture was heated to 80° C. for 72 h. Thereaction mixture was then cooled, diluted with dichloromethane (20 mL)and the organic phase was washed with saturated sodium bicarbonate (2×50mL) and brine (50 mL). The organic phase was then dried (magnesiumsulfate) and concentrated to give 6.5 g of a crude product. The crudeproduct was purified by chromatography on silica gel eluting with 3%methanol in dichloromethane to provide the title compound (1.4 g, 21%yield).

Preparation 15

Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]nonyl}piperidin-4-ylEster

Triethylamine hydrogen fluoride (376 μL, 2.3 mmol) was added to asolution of the product of Preparation 14 (1.3 g, 1.5 mmol) in THF (8mL) and the reaction mixture was stirred at ambient temperature. After 5h, the reaction was complete as determined by LCMS analysis. Thereaction mixture was then quenched with 1N NaOH until the pH was 14 andthen diluted with ethyl acetate (20 mL) and washed with 1N NaOH (20 mL)and brine (20 mL). The organic phase was then separated, dried overmagnesium sulfate, and concentrated to yield the title compound (1.1 g).

Example 3 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

A solution of the product of Preparation 15 (1.1 g, 1.5 mmol) wasflushed with nitrogen and palladium on carbon (10%, 110 mg) was added.The reaction mixture was stirred under hydrogen at balloon pressure.Analysis by LCMS showed that the reaction was completed after 9 h. Thereaction mixture was then filtered and concentrated to yield a yellowsolid. The solid was purified by preparative HPLC (5-30 over 60 min) toafford the title compound (510 mg). MS m/z: [M+H⁺] calcd for C₃₈H₄₈N₄O₅641.36. found 641. HPLC method 10-70: 3.207. [α]²⁰ _(D)=−23.6 (c=1.0mg/mL, water).

Example 3A Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

Alternatively, the title compound was prepared as follows:

(a) 9-Bromononanal

To a 100-mL round-bottomed flask equipped with a magnetic stirrer,addition funnel and temperature controller, under nitrogen, was added9-bromononanol (8.92 g, 40 mmol) and dichloromethane (30 mL). Theresulting mixture was cooled to 5° C. and a solution of sodiumbicarbonate (0.47 g, 5.6 mmol) and potassium bromide (0.48 g, 4 mmol) inwater (10 mL) was added. 2,2,6,6-Tetramethyl-1-piperidinyloxy freeradical (TEMPO) (63 mg, 0.4 mmol) was added and then a 10 to 13% bleachsolution (27 mL) was added dropwise through the addition funnel at arate such that the temperature was maintained at about 8° C. (+/−2° C.)with an ice cold bath (over about 40 min.). After addition of the bleachwas complete, the mixture was stirred for 30 min. while maintaining thetemperature at about 0° C. A solution of sodium bisulfite (1.54 g) inwater (10 mL) was added and the resulting mixture was stirred at roomtemperature for 30 min. The layers of the mixture were then separated,and the milky aqueous layer was extracted with dichloromethane (1×20mL). The combined dichloromethane layers were then washed with water(1×30 mL), dried (MgSO₄), filtered and concentrated under reducedpressure to afford the title intermediate (8.3 g, 94% yield), which wasused without further purification in the next step.

(b) 9-Bromo-1,1-dimethoxynonane

To a 100 mL round-bottomed flask was added 9-bromononanal (7.2 g, 32.5mmol), methanol (30 mL) and trimethylorthoformate (4 mL, 36.5 mmol). Asolution of 4 N hydrochloric acid in dioxane (0.2 mL, 0.8 mmol) wasadded and the resulting mixture was refluxed for 3 h. The reactionmixture was then cooled to room temperature and solid sodium bicarbonate(100 mg, 1.2 mmol) was added. The resulting mixture was concentrated toone-fourth its original volume under reduced pressure and then ethylacetate (50 mL) was added. The organic layer was washed with water (2×40mL), dried (MgSO₄), filtered and concentrated under reduced pressure toafford the title intermediate (8.44 g, (97% yield)) as a liquid, whichas used in the next step without further purification.

(c) Biphenyl-2-ylcarbamic Acid 1-(9,9-Dimethoxynonyl)piperidin-4-ylEster

To a 50 mL three-necked, round-bottomed flask was addedbiphenyl-2-ylcarbamic acid piperidin-4-yl ester (1 g, 3.38 mmol) andacetonitrile (10 mL) to form a slurry. To this slurry was added9-bromo-1,1-dimethoxynonane (1.1 g, 1.3 mmol) and triethylamine (0.57 g,4.1 mmol) and the resulting mixture was heated at 65° C. for 6 h (thereaction was monitored by HPLC until starting material is <5%). Thereaction mixture was then cooled to room temperature at which time themixture formed a thick slurry. Water (5 mL) was added and the mixturewas filtered to collect the solid on a coarse fritted glass filer. Thesolid was washed with pre-mixed solution of acetonitrile (10 mL) andwater (5 mL) and then with another pre-mixed solution of acetonitrile(10 mL) and water (2 mL). The resulting solid was air dried to affordthe title intermediate (1.37 g, 84%, purity >96% by LC, ¹H NMR) as awhite solid.

(d) Biphenyl-2-ylcarbamic Acid 1-(9-Oxononyl)piperidin-4-yl Ester

To a 500 mL round-bottomed flask with a magnetic stirrer was addedbiphenyl-2-ylcarbamic acid 1-(9,9-dimethoxynonyl)piperidin-4-yl ester(7.7 g, 15.9 mmol) and then acetonitrile (70 mL) and aqueous 1Mhydrochloric acid (70 mL). The resulting mixture was stirred at roomtemperature for 1 h and then dichloromethane (200 mL) was added. Thismixture was stirred for 15 min. and then the layers were separated. Theorganic layer was dried (MgSO₄), filtered and concentrated under reducedpressure to afford the title intermediate (6.8 g), which was used in thenext step without further purification.

(e) Biphenyl-2-ylcarbamic Acid1-(9-{Benzyl-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethyl]amino}nonyl)-piperidin-4-ylEster

To a 300 mL round-bottomed flask was added 5-[(R)-2-benzylamino1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one (5g, 9.73 mmol), dichloromethane (100 mL) and glacial acetic acid (0.6 mL,10 mmol). This mixture was cooled to 0° C. using an ice bath andbiphenyl-2-ylcarbamic acid 1-(9-oxononyl)piperidin-4-yl ester (4.6 g,9.73 mmol) was added with stirring. This mixture was stirred at 0° C.for 30 minutes and then sodium triacetoxyborohydride (6.15 g, 29 mmol)was added in portions over 15 minutes. The reaction mixture was stirredat 0° to 10° C. for 2 hours and then aqueous saturated sodiumbicarbonate solution (50 mL) was added and this mixture was stirred for15 minutes. The layers were then separated and the organic layer waswashed with 5% aqueous sodium chloride solution (50 mL), dried (MgSO₄),filtered and concentrated under reduced pressure to afford the titleintermediate (8.5 g, 80% purity by HPLC), which was used without furtherpurification.

(f) Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(tert-Butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster

To a 200 mL round-bottomed flask was added the intermediate from Step E(8.5 g, 9 mmol), ethanol (100 mL) and glacial acetic acid (0.54 mL, 18mmol) and this mixture was stirred until the solid dissolved. Thereaction mixture was purged with hydrogen for 5 min. and then 10%palladium on carbon (1.7 g) was added. This mixture was stirred at roomtemperature while hydrogen was slowly bubbling through the reactionmixture until >95% conversion was observed by HPLC (about 8-9 h). Themixture was then filtered through a Celite pad and the solvent wasremoved under reduced pressure. The residue was purified by silica gelchromatography (15 g silica/1 g crude) using 5% MeOH in DCM/0.5% NH₄OH(10×150 mL), 8% MeOH in DCM/0.5% NH₄OH (10×150 mL) and 10% MeOH inDCM/0.5% NH₄OH (10×150 mL). The appropriate fractions were combined andthe solvent was removed under reduced pressure while maintaining thetemperature <35° C. to give the title intermediate (4.05 g, 97% purity).

(g) Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster

To a 200 mL round-bottomed flask was added the intermediate from Step F(4.05 g, 5.36 mmol) and dichloromethane (80 mL) and the resultingmixture was stirred until the solid dissolved. Triethylaminetrihydrofluoride (2.6 mL, 16 mmol) was added and stirring was continuedunder nitrogen for 18 to 20 h. Methanol (20 mL) was added and thensaturated aqueous sodium bicarbonate (50 mL) was added slowly and themixture was stirred for 15 min. The layers were then separated and theorganic layer was washed with saturated aqueous sodium chloride solution(20 mL), dried (MgSO₄), filtered and concentrated under reduced pressureto afford the title compound (3.5 g, 98% purity by HPLC) as a yellowsolid.

Example 3B Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-Hydroxy-2-(8-Hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster Naphthalene-1,5-disulfonic Acid Salt

Biphenyl-2-ylcarbamic acid1-{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylester (1.0 g, 1.56 mmol, free base) was dissolved in methanol (10 mL;low water content). A solution of naphthalene-1,5-disulfonic acid (0.45g, 1.56 mmol) in methanol (5 mL; low water content) was added and thereaction mixture was stirred at 30° C. for two hours and then at roomtemperature overnight (18 h). The resulting thick slurry was filteredand the filtrate cake was washed with methanol (5 mL) and then dried togive the title compound (1.16 g, 80% yield) as off-white crystallinesolid.

Preparation 16

N-{2-Benzyloxy-5-[(R)-2-bromo-1-(tert-butyldimethylsilanyloxy)ethyl]phenyl}-formamide

(R)-2-Bromo-1-(3-formamido-4-benzyloxyphenyl)ethanol (9.9 g, 28 mmol)was dissolved in dimethylformamide (36 mL). Imidazole (2.3 g, 34 mmol)and tert-butyldimethylsilyl chloride (4.7 g, 31 mmol) were added. Thesolution was stirred under nitrogen atmosphere for 72 h. Additionalimidazole (0.39 g, 5.7 mmol) and tert-butyldimethylsilyl chloride (0.64g, 4.3 mmol) were added and the reaction was stirred for an additional20 h. The reaction mixture was then diluted with a mixture of isopropylacetate (53 mL) and hexanes (27 mL) and transferred to a separatoryfunnel. The organic layer was washed twice with a mixture of water (27mL) and saturated aqueous sodium chloride (27 mL) followed by a finalwash with saturated aqueous sodium chloride (27 mL). The organic layerwas dried over sodium sulfate. Silica gel (23.6 g) and hexanes (27 mL)were added and the suspension was stirred for 10 min. The solids wereremoved by filtration and the filtrate concentrated under vacuum. Theresidue was crystallized from hexanes (45 mL) to afford 8.85 g (19 mmol,68%) of the title compound as a solid. MS m/z: [M+H⁺] calcd forC₂₂H₃₀NO₃SiBr 464.1. found 464.2.

The starting material,(R)-2-bromo-1-(3-formamido-4-benzyloxyphenyl)ethanol, can be prepared asdescribed in U.S. Pat. No. 6,268,533 B1; or R. Hett et al., OrganicProcess Research and Development, 1998, 2:96-99; or using proceduressimilar to those described in Hong et al., Tetrahedron Lett., 1994,35:6631; or similar to those described in U.S. Pat. No. 5,495,054.

Preparation 17

Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(4-Benzyloxy-3-formylaminophenyl)-2-(tert-butyldimethylsilanyloxy)ethylamino]nonyl}piperidin-4-ylEster

The product of Preparation 16 (500 mg, 1.008 mmol) and sodium iodide(243 mg, 1.62 mmol) were stirred in tetrahydrofuran (0.5 mL) for 15 minat ambient temperature. The product of Preparation 11, (564 mg, 1.29mmol) and sodium bicarbonate (272 mg, 3.24 mmol) were then added and thereaction mixture was heated at 80° C. for 24 h. The reaction mixture wasthen allowed to cool. Water (2 mL) was then added and the mixture wasextracted with dichloromethane (2×2 mL). The combined organic extractswere washed with 1M hydrochloric acid (2×1 mL), dried (magnesiumsulfate) and the solvent was removed under reduced pressure. The cruderesidue was purified by flash chromatography (5-10%methanol/dichloromethane) to give the title compound (360 mg, 0.44 mmol,41% yield). HPLC (10-70) R_(t)=4.96; MS m/z: [M+H⁺] calcd for C₄₉H₆₈N₄O₅821.51. found 821.9.

Preparation 18

Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(4-Benzyloxy-3-formylaminophenyl)-2-hydroxyethylamino]nonyl}piperidin-4-ylEster

To a stirred solution of the product of Preparation 17 (360 mg, 0.44mmol) in tetrahydrofuran (2.2 mL) at ambient temperature was addedtriethylamine trihydrofluoride (108 μL, 0.66 mmol). The reaction mixturewas stirred for 24 h and then diluted with dichloromethane (5 mL) andwashed with 1M hydrochloric acid (2 mL) and saturated aqueous sodiumbicarbonate (2 mL). The organic phase was dried (magnesium sulfate) andthe solvent was removed under reduced pressure. The crude title compoundwas used directly in the next step without further purification. HPLC(10-70) R_(t)=4.6; MS m/z: [M+H⁺] calcd for C₄₃H₅₄N₄O₅ 707.43. found707.8.

Example 4 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-(3-Formylamino-4-hydroxyphenyl)-2-hydroxyethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

Palladium (10 wt. % (dry basis) on activated carbon) (124 mg) was addedto a stirred solution of the product of Preparation 18 (311 mg, 0.44mmol) in ethanol (4 mL) and the reaction mixture was placed under anatmosphere of hydrogen. After stirring for 12 h, the reaction mixturewas filtered through a pad of Celite, washed with methanol (2 mL) andthe solvent was removed under reduced pressure. The resulting residuewas purified by preparative HPLC to give the title compound (41 mg).HPLC (10-70) R_(t)=3.0; MS m/z: [M+H⁺] calcd for C₃₆H₄₈N₄O₅ 617.39.found 617.5.

Example 5 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)ethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

Palladium (10 wt. % (dry basis) on activated carbon) (80 mg) was addedto a stirred solution of the product of Example 3 (80 mg, 0.11 mmol) inethanol (1.1 mL) and the reaction mixture was placed under an atmosphereof hydrogen. The reaction mixture was stirred for 12 h, and thenfiltered through a pad of Celite, washed with methanol (2 mL) and thesolvent removed under reduced pressure. The crude material wasresubjected to the above conditions to ensure complete reaction. Theresulting residue was purified by preparative HPLC to yield the titlecompound (6 mg). HPLC (10-70) R_(t)=3.23; MS m/z: [M+H⁺] calcd forC₃₈H₅₀N₄O₅ 643.39. found 643.7.

Preparation 19

3-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]propionic Acid MethylEster

Methyl 3-bromopropionate (553 μL, 5.07 mmol) was added to a stirredsolution of the product of Preparation 8 (1.00 g, 3.38 mmol) and DIPEA(1.76 mL, 10.1 mmol) in acetonitrile (34 mL) at 50° C. and the reactionmixture was heated at 50° C. overnight. The solvent was then removedunder reduced pressure and the residue was dissolved in dichloromethane(30 mL). The resulting solution was washed with saturated aqueous sodiumbicarbonate solution (10 mL), dried (magnesium sulfate) and the solventwas removed under reduced pressure. The crude residue was purified bycolumn chromatography (5-10% MeOH/DCM) to give the title compound (905mg, 70%).

Preparation 20

3-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]propionic Acid

A stirred solution of the product of Preparation 19 (902 mg, 2.37 mmol)and lithium hydroxide (171 mg, 7.11 mmol) in 50% THF/H₂O (24 mL) washeated at 30° C. overnight, and then acidified with concentratedhydrochloric acid and lyophilized to give the title compound (˜100%yield, also contains LiCl salts).

Preparation 21

{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethyl-silanyloxy)ethylamino]pentyl}carbamicAcid tert-Butyl Ester

The product of Preparation 13 (600 mg, 1.23 mmol) andN-tert-butoxycarbonyl-1,5-diaminopentane (622 mg, 3.07 mmol) weredissolved in dimethyl sulfoxide (1.23 mL) and heated to 105° C. for 6 h.The reaction mixture was then cooled and diluted with ethyl acetate (10mL) and washed with saturated aqueous sodium bicarbonate solution (4mL). The organic phase was dried (magnesium sulfate) and the solvent wasremoved under reduced pressure. The crude residue was purified by columnchromatography (5-10% methanol/dichloromethane) to give the titlecompound (˜100% yield).

Preparation 22

5-[(R)-2-(5-Aminopentylamino)-1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

A solution of the product of Preparation 21 (800 mg, 1.31 mmol) intrifluoroacetic acid/dichloromethane (25%, 12 mL) was stirred at ambienttemperature for 1 hour. The solvent was then removed under reducedpressure and the crude residue was dissolved in dichloromethane (15 mL)and washed with 1N sodium hydroxide (8 mL). The organic phase wasseparated, dried (magnesium sulfate) and the solvent was removed underreduced pressure to give the title compound (509 mg, 81% yield over 2steps).

Preparation 23

Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]pentylcarbamoyl}ethyl)piperidin-4-ylEster

To the product of Preparation 20 (417 mg, 1.13 mmol) and HATU (430 mg,1.13 mmol) was added the product of Preparation 22 (458 mg, 0.90 mmol)in DMF (1.8 mL), followed by DIPEA (204 μL, 1.17 mmol). The reactionmixture was stirred at 50° C. for 12 h, and then the solvent was removedunder reduced pressure. The crude residue was dissolved indichloromethane (10 mL). The resulting solution was washed withsaturated aqueous sodium bicarbonate solution (4 mL), dried (magnesiumsulfate) and the solvent was removed under reduced pressure. The cruderesidue was purified by column chromatography (5-10%methanol/dichloromethane and 0.5% NH₄OH) to give the title compound (240mg, 31% yield).

Preparation 24

Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]pentylcarbamoyl}ethyl)piperidin-4-ylEster

To a stirred solution of the product of Preparation 23 (240 mg, 0.28mmol) in dichloromethane (2.8 mL) was added triethylaminetrihydrofluoride (91 μL, 0.56 mmol). The reaction mixture was stirredfor 10 h, and then diluted with dichloromethane (10 mL). The resultingsolution was then washed with saturated aqueous sodium bicarbonatesolution (5 mL), and then the organic phase was dried (magnesiumsulfate) and the solvent was removed under reduced pressure to give thetitle compound (209 mg, 100% yield).

Example 6 Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylcarbamoyl}ethyl)piperidin-4-ylEster, Ditrifluoroacetate

To a stirred solution of the product of Preparation 24 (209 mg, 0.28mmol) in ethanol (2.8 mL) was added palladium (10 wt. % (dry basis) onactivated carbon) (81 mg) and the reaction mixture was placed under anatmosphere of hydrogen and stirred overnight. The reaction mixture wasthen filtered and solvent was removed under reduced pressure. The cruderesidue was purified by preparative HPLC to give the title compound (58mg). HPLC (10-70) R_(t)=2.30; MS m/z: [M+H⁺] calcd for C₃₇H₄₅N₅O₆656.34. found 656.6; [α]²⁰ _(D)=−6.5 (c=11.0 mg/mL, water).

Example 6A Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylcarbamoyl}ethyl)piperidin-4-ylEster

Alternatively, the title compound can be prepared as follows:

(a) 5-Chloropentanal

To a 2 L three-necked round-bottomed flask, equipped with a magneticstirrer, addition funnel and temperature controller under nitrogen, wasadded 5-chloropentanol (53 g, 0.433 mol) and dichloromethane (300 mL).This mixture was cooled to 5° C. and a solution of sodium bicarbonate (5g, 0.059 mol) and potassium bromide (5.1 g, 0.043 mol) in water (225 mL)was added. 2,2,6,6-Tetramethyl-1-piperidinyloxy free radical (TEMPO) (63mg, 0.4 mmol) was added and then a 10 to 13% bleach solution (275 mL)was added dropwise through the addition funnel at a rate such that thetemperature was maintained at about 8° C. (+/−2° C.) with an ice coldbath (over about 45 min.). After addition of the bleach was complete,the mixture was stirred for 30 min. while maintaining the temperature atabout 5° C. A solution of sodium bisulfite (4 g) in water (30 mL) wasadded and the resulting mixture was stirred at room temperature for 30min. The layers of the mixture were then separated, and the aqueouslayer was extracted with dichloromethane (1×50 mL). The combineddichloromethane layers were then washed with water (1×50 mL), dried(MgSO₄), filtered and concentrated under reduced pressure to afford thetitle compound (53 g). The product was distilled at 65° C./8 torr toafford the title compound (31.16 g) as an orange oil (GC purity was 70to 80%).

The product was further purified by adding the crude material (4 g) to amixture of ethanol (920 mL), ethyl acetate (12 mL) and water (4 mL).Sodium bisulfite (4 g) was added and the mixture was heated to refluxfor 4 h and then cooled to room temperature and stirred for 14 h at roomtemperature to form a very thick slurry. The solids were filtered on acoarse fritted filter, washed with the solvent mixture (5 mL) and thesolids were dried on the filter to afford 8.4 g of the bisulfite adduct.This material was then added to MTBE (20 mL) and aqueous 1 N sodiumhydroxide (45 mL) was added with vigorous stirring. The resultingbiphasic mixture was stirred vigorously until all the solids haddissolved (about 15 min) and then the layers were separated. The aqueouslayer was extracted with MTBE (20 mL) and the combined MTBE layers weredried (MgSO₄), filtered and concentrated to afford 3.46 g of the titlecompound as a colorless liquid (GC purity >90%).

(b)5-[(R)-2-[Benzyl-(5-chloropentyl)amino]-1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

To a 1 L three-necked round-bottomed flask was added the product ofPreparation 28 (48.4 g, 94 mmol), dichloromethane (400 mL) and glacialacetic acid (11.3 mL). This mixture was stirred at 0° C. (ice bath) andthe product from step (a) (12.5 g, 103.6 mmol) was added and stirringwas continued for 15 min. Sodium triacetoxyborohydride (59.8 g, 282mmol) was then added in portions over a 15 min. period and the resultingmixture was stirred at 0° C. to 10° C. for 2 h. Aqueous saturated sodiumbicarbonate solution (200 mL) was then added slowly (gas evolution) andstirring was continued for 15 min. The pH of the solution was thenadjusted with solid sodium carbonate to a pH of about 9 and the layerswere separated. The organic layer was washed with aqueous 5% sodiumchloride solution (200 mL), dried (MgSO₄), filtered and concentratedunder reduced pressure to afford the title compound (53 g).

(c)5-[(R)-2-[(5-N,N-Diformylaminopentyl)benzylamino]-1-(tert-butyldimethyl-silanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

To a stirred solution of the product of step (b) (26.5 g, 42.8 mmol) in1-methyl-2-pyrrolidinone (175 mL) was added sodium diformylamide (6.1 g,64.2 mmol) and sodium iodide (2.13 g, 14.3 mmol). The reaction flask wasflushed with nitrogen and then the mixture was heated at 65° C. for 8 h.The mixture was then cooled to room temperature and water (300 mL) andethyl acetate (100 mL) were added. This mixture was stirred for 10 min.and then the layers were separated. The aqueous layer was extracted withethyl acetate (150 mL) and the combined organic layers were washed withwater (300 mL), aqueous 50% brine solution (300 mL), water (300 mL),dried (MgSO₄) filtered and concentrate to afford the title compound(23.3 g).

(d)5-[(R)-2-[(5-Aminopentyl)benzylamino]-1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

To a stirred solution of the product from step (c) (10.5 g, 16 mmol) inmethanol (75 mL) was added p-toluenesulfonic acid (7.42 g. 39 mmol). Theresulting mixture was heated at 40° C. for 15 h and then concentratedunder reduced pressure to about half its volume. Methanol (70 mL) wasadded and the mixture was heated at 50° C. for 2 h and then concentratedunder reduced pressure. Water (100 mL), methanol (50 mL) and MTBE (100mL) were added and this mixture was stirred for 15 min and then thelayers were separated. To the aqueous layer was added aqueous 1 N sodiumhydroxide (45 mL) and MTBE (100 mL), and this mixture was stirred for 15min. The layers were then separated and the aqueous layer was extractedwith MTBE (100 mL). The combined MTBE layers were dried (MgSO₄),filtered and concentrated to afford the title compound as a yellow oil(7.3 g). This material contained about 13% (by HPLC) of thecorresponding des-tert-butyldimethylsilyl compound.

(e) 3-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]propionic Acid

To a solution of the product of Preparation 8 (50 g, 67.6 mmol) indichloromethane (500 mL) was added acrylic acid (15.05 mL, 100 mmol).The resulting mixture was heated at 50° C. under reflux for 18 h andthen the solvent was removed. Methanol (600 mL) was added and thismixture was heated at 75° C. for 2 h and then cooled to room temperatureto form a thick slurry. The solid was collected by filtration, washedwith methanol (50 mL) and air dried to afford the title compound (61g, >96% purity) as white powder.

(f) Biphenyl-2-ylcarbamic Acid1-[2-(5-{(Benzyl-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethyl]amino}-pentylcarbamoyl)ethyl]piperidin-4-ylEster

A mixture of the product of step (e) (3.68 g, 10 mmol) andN,N-dimethylformamide (50 mL) was heated at 60° C. until the solidcompletely dissolved and then cooled to room temperature. The product ofstep (d) (6 g, 10 mmol) and diisopropylethylamine (3.5 mL) was added andthe reaction mixture was cooled to 0° C. PyBOP (6.25 g, 12 mmol) wasadded in one portion and the reaction mixture was stirred at 0° C. toroom temperature for 2 hours. The reaction mixture was then poured intocold water (500 mL) with stirring and the pH of the resulting mixturewas adjusted to about 2 using aqueous 1 M hydrochloric acid. Thismixture was stirred for 15 min and then filtered to collect the solid,which was washed with water (100 mL) and dried to afford the titlecompound (8.7 g, HPLC purity >95%) as an off-white solid.

(g) Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylcarbamoyl}ethyl)piperidin-4-ylEster

The product of step (f) can be deprotected using essentially the sameprocedures as those described in Preparation 24 and Example 6 to affordthe title compound.

Preparation 25

2-(N-Benzyloxycarbonyl-N-methylamino)ethanal (a)2-(N-Benzyloxycarbonyl-N-methylamino)ethanol

Benzyl chloroformate (19 g, 111.1 mmol) in THF (20 mL) was addeddropwise over 15 min to a stirred solution of 2-(methylamino)ethanol (10g, 133.3 mmol) in THF (100 mL) and aqueous sodium carbonate (100 mL) at0° C. The reaction mixture was stirred at 0° C. for 12 h and thenextracted with EtOAc (2×200 mL). The organic layer was washed withaqueous sodium carbonate (200 mL) and dried (potassium carbonate) andsolvent was removed under reduced pressure to give the title compound(22.5 g, 97% yield).

(b) 2-(N-Benzyloxycarbonyl-N-methylamino)ethanal

DMSO (71 mL, 1 mol) and DIPEA (87.1 mL, 0.5 mol) were added to a stirredsolution of the product of step (a) (20.9 g, 0.1 mol) in dichloromethane(200 mL) at −10° C. The reaction mixture was stirred at −10° C. for 15min and then sulfur trioxide pyridine complex (79.6 g, 0.5 mol) wasadded and the resulting mixture was stirred for 1 hour. The reactionmixture was quenched with addition of 1M hydrochloric acid (200 mL). Theorganic layer was separated and washed with saturated aqueous sodiumbicarbonate (100 mL), brine (100 mL), dried (potassium carbonate) andsolvent removed under reduced pressure to give the title compound (20.7g, ˜100% yield).

Preparation 26

Biphenyl-2-ylcarbamic Acid 1-[2-(methylamino)ethyl]piperidin-4-yl Ester

To a stirred solution of the product of Preparation 25 (20.7 g, 100mmol) and the product of Preparation 8 (25 g, 84.7 mmol) in MeOH (200mL) was added sodium triacetoxyborohydride (21.2 g, 100 mmol). Thereaction mixture was stirred for 12 h at ambient temperature and then itwas quenched with 2M hydrochloric acid and the solvent was removed underreduced pressure. The residue was dissolved in ethyl acetate (200 mL)and washed with saturated aqueous sodium bicarbonate solution (100 mL)and brine (50 mL), and then dried (magnesium sulfate) and the solventwas removed under reduced pressure. The crude residue was purified bycolumn chromatography (50-90% EtOAc/hexanes) to givebiphenyl-2-ylcarbamic acid1-[2-(benzyloxycarbonyl-methylamino)ethyl]piperidin-4-yl ester as anoil.

The oil was dissolved in methanol (100 mL) and palladium (10 wt. % (drybasis) on activated carbon) (5 g) was added. The reaction mixture wasstirred under hydrogen (30 psi) for 12 h and then filtered throughCelite, which was washed with methanol, and solvent was evaporated togive the title compound (13.2 g, 44% yield).

Preparation 27

Biphenyl-2-ylcarbamic Acid1-{2-[(6-Bromohexanoyl)methylamino]ethyl}piperidin-4-yl Ester

6-Bromohexanoyl chloride (3.23 mL, 21.1 mmol) was added to a stirredsolution of the product of Preparation 26 (6.2 g, 17.6 mmol) and DIPEA(6.13 mL, 35.2 mmol) in dichloroethane (170 mL). The reaction mixturewas stirred for 1 hour and it was then diluted with EtOAc (250 mL) andwashed with saturated aqueous sodium bicarbonate solution (2×200 mL) andbrine (200 mL), and then dried (magnesium sulfate). The solvent wasremoved under reduced pressure to give the title compound (6.6 g, 73%yield).

Preparation 28

8-Benzyloxy-5-[(R)-2-(N-benzylamino)-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one

A stirred solution of the product of Preparation 13 (1.00 g, 2.05 mmol)and benzylamine (493 μL, 4.51 mmol) in DMSO (1.7 mL) was heated at 105°C. for 4 h. The reaction mixture was allowed to cool and was thendiluted with EtOAc (10 mL) and the organic layer was washed withsaturated aqueous ammonium chloride solution (5 mL) and 1N sodiumhydroxide (5 mL), dried (MgSO₄) and solvent removed under reducedpressure. The crude residue was purified by column chromatography (50%EtOAc/hexanes) to give the title compound (700 mg, 67%). MS m/z: [M+H⁺]calcd for C₃₁H₃₈N₂O₃Si 515.27. found 515.5.

Preparation 29

Biphenyl-2-ylcarbamic Acid1-{2-[(6-{Benzyl-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethyl]amino}hexanoyl)-methylamino]ethyl}piperidin-4-ylEster

To a stirred solution of the product of Preparation 28 (807 mg, 1.57mmol) and DIPEA (819 μL, 4.7 mmol) in acetonitrile (3.14 mL) was addedthe product of Preparation 27 (995 mg, 1.88 mmol). The reaction mixturewas heated to 80° C. for 24 h. The solvent was removed under reducedpressure and the residue was dissolved in EtOAc (10 mL) and then washedwith saturated aqueous sodium bicarbonate solution (5 mL), dried(magnesium sulfate), and the solvent removed under reduced pressure. Thecrude material was purified by column chromatography (4-6% MeOH/DCM) toobtain the title compound (452 mg, 30% yield).

Preparation 30

Biphenyl-2-ylcarbamic Acid1-{2-[(6-{Benzyl-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethyl]amino}hexanoyl)methylamino]ethyl}piperidin-4-ylEster

To a stirred solution of the product of Preparation 29 (452 mg, 0.47mmol) in dichloromethane (4.7 mL) was added triethylaminetrihydrofluoride (116 μL, 0.71 mmol). The reaction mixture was stirredfor 10 h. and then it was diluted with dichloromethane (10 mL) andwashed with saturated aqueous sodium bicarbonate solution (5 mL). Theorganic phase was then dried (MgSO₄) and the solvent was removed underreduced pressure to give the title compound (100% yield).

Example 7 Biphenyl-2-ylcarbamic Acid1-[2-({6-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]hexanoyl}methylamino)ethyl]piperidin-4-ylEster Ditrifluoroacetate

To a stirred solution of the product of Preparation 30 (400 mg, 0.47mmol) in ethanol (4.7 mL) was added palladium (10 wt. % (dry basis) onactivated carbon) (160 mg) and the reaction mixture was placed under anatmosphere of hydrogen and stirred overnight. The reaction mixture wasthen filtered and solvent was removed under reduced pressure. The cruderesidue was purified by preparative HPLC to give the title compound (73mg). HPLC (10-70) R_(t)=2.33; MS m/z: [M+H⁺] calcd for C₃₈H₄₇N₅O₆670.36. found 670. [α]²⁰ _(D)=−9.4 (c=1.0 mg/mL, water).

Preparation 31

Biphenyl-2-ylcarbamic Acid1-[2-(4-(Aminomethyl)phenylcarbamoyl)-ethyl]piperidin-4-yl Ester

To a stirred solution of 4-(N-tert-butoxycarbonylaminomethyl)aniline(756 mg, 3.4 mmol), the product of Preparation 20 (1.5 g, 4.08 mmol) andHATU (1.55 g, 4.08 mmol) in DMF (6.8 mL) was added DIPEA (770 μL, 4.42mmol). The reaction mixture was stirred at 50° C. overnight and then thesolvent was removed under reduced pressure. The resulting residue wasdissolved in dichloromethane (20 mL) and washed with saturated aqueoussodium bicarbonate solution (10 mL). The organic phase was then dried(magnesium sulfate) and the solvent was removed under reduced pressure.The crude product was purified by flash chromatography (5-10% MeOH/DCM)to give a solid, which was dissolved in TFA/DCM (25%, 30 mL) and stirredat room temperature for 2 h. The solvent was then removed under reducedpressure and the crude residue was dissolved in dichloromethane (30 mL)and washed with 1N sodium hydroxide (15 mL). The organic phase wasseparated, dried (magnesium sulfate) and the solvent was removed underreduced pressure to give the title compound (1.5 g, 94% over 2 steps).

Preparation 32

Biphenyl-2-ylcarbamic Acid1-[2-(4-([(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]methyl)phenylcarbamoyl)ethyl]-piperidin-4-ylEster

A solution of the product of Preparation 31 (489 mg, 1.04 mmol), theproduct of Preparation 13 (610 mg, 1.25 mmol), sodium bicarbonate (262mg, 3.12 mmol) and sodium iodide (203 mg, 1.35 mmol) in THF (0.52 mL)were heated at 80° C. for 12 h. The reaction mixture was diluted withdichloromethane (10 mL) and washed with saturated aqueous sodiumbicarbonate solution (5 mL). The organic phase was dried (MgSO₄) and thesolvent was removed under reduced pressure. The crude residue waspurified by flash chromatography (10% MeOH/DCM) to give the titlecompound as a solid (687 mg, 77% yield).

Preparation 33

Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-ylEster

To a stirred solution of the product of Preparation 32 (687 mg, 0.8mmol) in dichloromethane (8 mL) was added triethylamine trihydrofluoride(261 μL, 1.6 mmol). The reaction mixture was stirred for 10 h and thenwas diluted with dichloromethane (20 mL) and washed with saturatedaqueous sodium bicarbonate solution (10 mL). The organic phase was thendried (magnesium sulfate) and the solvent was removed under reducedpressure to yield the title compound (500 mg, 81% yield).

Example 8 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-ylEster Ditrifluoroacetate

To a stirred solution of the product of Preparation 33 (500 mg, 0.65mmol) in ethanol (6.5 mL) was added palladium (10 wt. % (dry basis) onactivated carbon) (200 mg) and the reaction mixture was placed under ahydrogen atmosphere and stirred overnight. The reaction mixture was thenfiltered and the solvent was removed under reduced pressure. The cruderesidue was purified by preparative HPLC to give the title compound (81mg, 2 TFA salt). HPLC (10-70) R_(t)=2.41; MS m/z: [M+H⁺] calcd forC₃₉H₄₁N₅O₆ 676.32. found 676.5.

Preparation 34

Biphenyl-2-ylcarbamic Acid1-(2-tert-Butoxycarbonylaminoethyl)piperidin-4-yl Ester

To a stirred solution of the product of Preparation 8 (2.00 g, 6.76mmol) and DIPEA (3.54 mL, 20.3 mmol) in acetonitrile (67.6 mL) at 50° C.was added 2-tert-butoxycarbonylaminoethyl bromide (1.82 g, 8.11 mmol)and the reaction mixture was heated at 50° C. overnight. The solvent wasthen removed under reduced pressure and the residue was dissolved indichloromethane (60 mL) and washed with saturated aqueous sodiumbicarbonate solution (30 mL). The organic phase was dried (magnesiumsulfate) and the solvent was removed under reduced pressure. The cruderesidue was purified by column chromatography (5% MeOH/DCM) to yield thetitle compound as a solid (2.32 g, 78% yield).

Preparation 35

Biphenyl-2-ylcarbamic Acid 1-(2-Aminoethyl)piperidin-4-yl Ester

The product of Preparation 34 was dissolved in TFA/DCM (25%, 52 mL) andstirred at room temperature for 2 h. The solvent was then removed underreduced pressure and the crude residue dissolved in dichloromethane (30mL) and washed with 1N sodium hydroxide (15 mL). The organic phase wasseparated, dried (magnesium sulfate) and the solvent was removed underreduced pressure to give the title compound (1.61 g, 90% yield).

Preparation 36

Biphenyl-2-ylcarbamic Acid1-[2-(4-Aminomethylbenzoylamino)ethyl]piperidin-4-yl Ester

To a stirred solution of the product of Preparation 35 (339 mg, 1 mmol),4-(tert-butoxycarbonylaminomethyl)benzoic acid (301 mg, 1.2 mmol) andHATU (456 mg, 1.2 mmol) in DMF (2 mL) was added DIPEA (226 μL, 1.3mmol). The reaction mixture was stirred at room temperature overnightand then the solvent was removed under reduced pressure. The resultingresidue was dissolved in dichloromethane (20 mL) and washed withsaturated aqueous sodium bicarbonate solution (10 mL). The organic phasewas dried (magnesium sulfate) and the solvent was removed under reducedpressure. The crude product was dissolved in TFA/DCM (25%, 10 mL) andthis mixture was stirred at room temperature for 2 h. The solvent wasremoved under reduced pressure and the crude residue was dissolved indichloromethane (15 mL) and washed with 1N sodium hydroxide (5 mL). Theorganic phase was separated, dried (magnesium sulfate) and the solventwas removed under reduced pressure to afford the title compound (472 mg,˜100% over 2 steps).

Preparation 37

Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]methyl}benzoylamino)ethyl]-piperidin-4-ylEster

A solution of the product of Preparation 36 (520 mg, 1.1 mmol), theproduct of Preparation 13 (634 mg, 1.3 mmol), sodium bicarbonate (277mg, 3.3 mmol) and sodium iodide (215 mg, 1.43 mmol) in THF (0.55 mL) washeated at 80° C. for 12 h. The reaction mixture was then diluted withdichloromethane (10 mL) and washed with saturated aqueous sodiumbicarbonate solution (5 mL). The organic phase was then dried (magnesiumsulfate) and the solvent was removed under reduced pressure. The cruderesidue was purified by flash chromatography (5-10% MeOH/DCM) to givethe title compound as a solid (316 mg, 33% yield).

Preparation 38

Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]methyl}benzoylamino)ethyl]piperidin-4-ylEster

To a stirred solution of the product of Preparation 37 (316 mg, 0.36mmol) in dichloromethane (3.6 mL) was added triethylaminetrihydrofluoride (117 μL, 0.72 mmol). The reaction mixture was stirredfor 10 h and was then diluted with dichloromethane (10 mL) and washedwith saturated aqueous sodium bicarbonate solution (5 mL). The organicphase was dried (MgSO₄) and the solvent was removed under reducedpressure to give the title compound, which was used directly in the nextstep (100% yield).

Example 9 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzoylamino)ethyl]piperidin-4-ylEster Ditrifluoroacetate

To a stirred solution of the product of Preparation 38 (275 mg, 0.36mmol) in ethanol (3.6 mL) was added palladium (10 wt. % (dry basis) onactivated carbon) (275 mg) and the reaction mixture was placed under ahydrogen atmosphere and stirred overnight. The reaction mixture was thenfiltered and the solvent was removed under reduced pressure. The cruderesidue was purified by preparative HPLC to yield the title compound (6mg, 2 TFA salt). HPLC (10-70) R_(t)=2.26; MS m/z: [M+H⁺] calcd forC₃₉H₄₁N₅O₆ 676.32. found 676.5.

Preparation 39

Biphenyl-2-ylcarbamic Acid 1-(2-Aminoethyl)piperidin-4-yl Ester

2-tert-Butoxycarbonylaminoethyl bromide (1.22 g, 5.44 mmol) was added toa solution of the product of Preparation 8 (1.46 g, 4.95 mmol) anddiisopropylethylamine (1.03 mL, 5.94 mmol) in acetonitrile (24 mL). Thereaction mixture was stirred at 65° C. for 12 hours, at which time MSanalysis showed that the reaction was completed. The reaction mixturewas concentrated to dryness and then dichloromethane (10 mL) was added.Trifluoroacetic acid was added to this mixture and the mixture wasstirred at room temperature for 4 hours, at which time MS analysisshowed that the reaction was complete. The mixture was then concentratedto half its volume and 1N sodium hydroxide was added to the solutionuntil the pH was adjusted to 14. The organic layer was washed withbrine, then dried over magnesium sulfate and filtered. The filtrate wasconcentrated to give 1.6 g of the title compound as a solid. MS m/z:[M+H⁺] calcd for C₂₀H₂₅N₃O₂ 340.2. found 340.

Preparation 40

5-[(R)-2-(5-Aminopentylamino)-1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

N-tert-butoxycarbonyl-1,5-diaminopentane (1.04 g, 5.12 mmol) was addedto a solution of the product of Preparation 13 (1.00 g, 2.05 mmol) indimethyl sulfoxide (2 mL). The solution was stirred at 75° C. for 12hours, at which time LCMS analysis showed that the reaction wascomplete. The reaction mixture was then concentrated under vacuum todryness. To the residue was added dichloromethane (2 mL) andtrifluoroacetic acid (1 mL) was then added. The solution was stirred atroom temperature for about 3 hours, at which time MS analysis showedthat the reaction was complete. The solution was concentrated to halfits volume and 1N sodium hydroxide was added until the pH was adjustedto 14. The organic layer was collected, washed with brine, dried overmagnesium sulfate and then concentrated to yield 782 mg of the titlecompound as an oil. MS m/z: [M+H⁺] calcd for C₂₉H₄₃N₃O₃Si 510.8. found510.

Preparation 41

Biphenyl-2-ylcarbamic Acid1-[2-(3-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]pentyl}-ureido)ethyl]piperidin-4-ylEster

Carbonyl diimidazole (127 mg, 0.78 mmol) was added to a solution of theproduct of Preparation 39 (266 mg, 0.78 mmol) in dimethyl formamide (4mL) and the resulting mixture was stirred at room temperature for 3hours. After 3 hours, the product of Preparation 40 (399 mg, 0.78 mmol)was added to the reaction mixture and this mixture was stirred for 12hours at room temperature, at which time LCMS analysis determined thatthe reaction was complete. The reaction mixture was concentrated invacuo and the residue was diluted with ethyl acetate (5 mL). The organiclayer was washed two times with saturated sodium bicarbonate (5 mL) andthen brine (5 mL). The organic layer was dried over magnesium sulfate,filtered and then concentrated to afford 597 mg of the title compound asa solid which was used without further purification. MS m/z: [M+H⁺]calcd for C₅₀H₆₆N₆O₆Si 875.5. found 875.

Preparation 42

Biphenyl-2-ylcarbamic Acid1-[2-(3-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]pentyl}ureido)ethyl]piperidin-4-ylEster

Triethylamine trihydrofluoride (0.16 mL, 1.02 mmol) was added to asolution of the product of Preparation 41 (597 mg, 0.68 mmol) intetrahydrofuran (3.4 mL) and this mixture was stirred at roomtemperature for about 12 hours, at which time the reaction wasdetermined to be completed by MS analysis. The reaction mixture wasdiluted with ethyl acetate (5 mL) and this mixture was washed with 1Nsodium hydroxide (5 mL), brine, dried over magnesium sulfate andconcentrated to give 417 mg of the title compound as a solid. MS m/z:[M+H⁺] calcd for C₄₄H₅₁N₆O₆ 760.4. found 760.

Example 10 Biphenyl-2-ylcarbamic Acid1-[2-(3-{5-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentyl}ureido)ethyl]piperidin-4-ylEster Ditrifluoroacetate

A solution of the product of Preparation 42 (417 mg, 0.55 mmol) inethanol (3 mL) was purged with nitrogen for about 10 minutes. Palladium(10 wt. % (dry basis) on activated carbon) (200 mg) was added and thesolution was flushed again with nitrogen for about 10 minutes. The flaskwas purged under vacuum and then filled with nitrogen three times andthen a hydrogen-filled balloon was placed over the flask. The reactionmixture was stirred under hydrogen for 12 hours, at which time thereaction was determined to be complete by MS analysis. The reactionmixture was then filtered and the organic filtrate concentrated andpurified by HPLC (10-35% over 60 minute) to give 146 mg of the titlecompound as a powder. MS m/z: [M+H⁺] calcd for C₃₇H₄₆N₆O₆ 671.4. found670. HPLC (10-70) R_(t)=2.6 minutes.

Example 11 Biphenyl-2-ylcarbamic Acid1-[3-(3-{5-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentyl}ureido)propyl]piperidin-4-ylEster Ditrifluoroacetate

Using the method described above in Preparations 39-42 and Example 10,and substituting 3-tert-Butoxycarbonylaminoprop-1-yl bromide for2-tert-butoxycarbonylaminoethyl bromide in Preparation 39, the titlecompound was prepared. MS m/z: [M+H⁺] calcd for C₃₈H₄₈N₆O₆ 685.4. found684. HPLC (10-70) R_(t)=2.6 minutes.

Preparation 43

6-(2-Bromo-(R)-1-tert-butyldimethylsilyloxy)ethyl-2,2-dimethyl-1,3-benzodioxan(a) 6-Bromo-2,2-dimethyl-4H-benzo[1,3]dioxine

To 5-bromo-2-hydroxybenzyl alcohol (93 g, 0.46 mol, available fromSigma-Aldrich) in 2.0 L of 2,2-dimethoxypropane was added 700 mL ofacetone, followed by zinc chloride (170 g). After stirring for 18 hours,1.0 M aqueous sodium hydroxide was added until the aqueous phase wasbasic. Diethyl ether (1.5 L) was added to the slurry and the organicphase was decanted into a separatory funnel. The organic phase waswashed with brine, dried over Na₂SO₄, filtered and concentrated underreduced pressure to give the title compound as an oil.

(b) 6-Acetyl-2,2-dimethyl-4H-benzo[1,3]dioxine

To the product of step (a) (110 g, 0.46 mol) in 1.0 L of THF at −78° C.was added 236 mL (0.51 mol) of 2.14 M n-butyllithium in hexanes via adropping funnel. After 30 minutes, N-methyl-N-methoxy acetamide (71 g,0.69 mol, available from TCI) was added. After 2 hours, the reactionmixture was quenched with water, diluted with 2.0 L of 1.0 M aqueousphosphate buffer (pH=7.0) and extracted once with diethyl ether. Thediethyl ether phase was washed once with brine, dried over Na₂SO₄,filtered and concentrated under reduced pressure to give a light orangeoil. The oil was dissolved in a minimum volume of ethyl acetate, dilutedwith hexanes, and to give the title compound as a crystalline solid.

(c) 6-Bromoacetyl-2,2-dimethyl-4H-benzo[1,3]dioxine

To the product of step (b) (23.4 g, 0.113 mol) in 600 mL of THF at −78°C. was added 135 mL of 1.0 M sodium hexamethyldisilazane in THF(Sigma-Aldrich). After 1 hour, trimethylsilyl chloride (15.8 mL, 0.124mol) was added. After another 30 minutes, bromine (5.82 mL, 0.113 mol)was added. After 10 minutes, the reaction was quenched by diluting thereaction mixture with diethyl ether and pouring it onto 500 mL of 5%aqueous Na₂SO₃ premixed with 500 mL of 5% aqueous NaHCO₃. The phaseswere separated and the organic phase was washed with brine, dried overNa₂SO₄, filtered and concentrated under reduced pressure to give thetitle compound as an oil that solidified upon storage in the freezer.

(d) (R)-2-Bromo-1-(2,2-dimethyl-4H-benzo[1,3]dioxin-6-yl)ethanol

To the product of step (c) (10 g, 35.1 mmol) in 100 mL of THF was addedthe solid catalyst of Preparation 13, step (c) (1) (0.97 g, 3.5 mmol).The solution was cooled to between −20° C. and −10° C. and BH₃-THF (35mL, 35 mmol) diluted with 50 mL THF was added dropwise via a droppingfunnel. After the addition was complete, the reaction mixture wasallowed to warm to ambient temperature. After 30 minutes, the reactionmixture was quenched by slow addition of 50 mL of methanol and thenconcentrated to a thick oil. The oil was purified by silica gelchromatography eluted with 1:2 ethyl acetate/hexanes. The fractions werecombined and concentrated to give the title compound as an off-whitesolid.

(e)[(R)-2-Bromo-1-(2,2-dimethyl-4H-benzo[1,3]dioxin-6-yl)ethoxy]-tert-butyldimethylsilane

To the product of step (d) (10 g, 34.8 mmol) and imidazole (4.7 g, 69.7mmol) dissolved in 100 mL DMF was added tert-butyldimethylsilyl chloride(5.78 g, 38.3 mmol). The reaction mixture was stirred for 18 hours. Thereaction mixture was then partitioned between 200 mL of saturated sodiumchloride and 200 mL of diethyl ether. The aqueous layer was extractedwith 200 mL of diethyl ether. The organic layers were then combined,washed with saturated sodium chloride (3×100 mL), dried over MgSO₄ andconcentrated. The product was purified by silica gel chromatography,eluting with hexanes followed by 5% ethyl acetate in hexanes. Thedesired fractions were combined and concentrated to give the titlecompound as an oil.

Preparation 44

Biphenyl-2-ylcarbamic Acid1-{9-[2-(tert-Butyldimethylsilanyloxy)-2-(2,2-dimethyl-4H-benzo[1,3]dioxin-6-yl)ethylamino]nonyl}piperidin-4-ylEster

The product of Preparation 43 (802 mg, 2.00 mmol) and sodium iodide (300mg, 2.00 mmol) were stirred in tetrahydrofuran (0.77 mL) for 15 min atambient temperature. The product of Preparation 11 (675 mg, 1.54 mmol)and sodium bicarbonate (388 mg, 4.62 mmol) were added and the reactionmixture was heated at 80° C. for 24 h. The reaction mixture was thencooled and water (2 mL) was added. The mixture was then extracted withdichloromethane (2×2 mL). The combined organic extracts were dried(magnesium sulfate) and the solvent was removed under reduced pressure.The crude residue was purified by flash chromatography (5-10%methanol/dichloromethane) to give the title compound as a solid (798 mg,1.05 mmol, 60% yield). MS m/z: [M+H⁺] calcd for C₄₅H₆₇N₃O₅Si 758.5.found 758.6.

Preparation 45

Biphenyl-2-ylcarbamic Acid1-{9-[2-(2,2-Dimethyl-4H-benzo[1,3]dioxin-6-yl)-2-hydroxyethylamino]nonyl}piperidin-4-ylEster

Triethylamine trihydrofluoride (342 μL, 2.10 mmol) was added to astirred solution of the Product of Preparation 44 (798 mg, 1.05 mmol) indichloromethane (10.5 mL) at ambient temperature. The reaction mixturewas stirred for 24 h and it was then diluted with dichloromethane (20mL) and washed with saturated aqueous sodium bicarbonate (15 mL). Theorganic layer was dried (magnesium sulfate) and the solvent was removedunder reduced pressure. The crude title compound was isolated as an oil(659 mg, 1.02 mmol), which was used in the next step without furtherpurification. MS m/z: [M+H⁺] calcd for C₃₉H₅₃N₃O₅ 644.4. found 644.8.

Example 12 Biphenyl-2-ylcarbamic Acid1-{9-[(R)-2-Hydroxy-2-(4-hydroxy-3-hydroxymethylphenyl)ethylamino]nonyl}piperidin-4-ylEster Ditrifluoroacetate

Trifluoroacetic acid (2.80 mL) was added to a stirred solution of theproduct of Preparation 45 (600 mg, 0.93 mmol) in THF/H₂O (14 mL, 1:1)and the reaction mixture was stirred for 2 h at ambient temperature. Thereaction mixture was concentrated under reduced pressure and dissolvedin 20% MeCN/H₂O then purified by preparative HPLC to yield the titlecompound (200 mg, 2TFA salt). HPLC (10-70) R_(t)=2.76; MS m/z: [M+H⁺]calcd for C₃₆H₄₉N₃O₅ 604.4. found 604.8.

Preparation 46

1-[1-(9-Benzylaminononyl)piperidin-4-yl]-3-biphenyl-2-ylurea

N-Benzylamine (0.903 ml, 8.30 mmol) was added to a solution of theproduct of Preparation 4 (2.40 g, 5.52 mmol) in methanol (25 mL) and theresulting mixture was stirred at ambient temperature. After 10 min,sodium triacetoxyborohydride (1.75 g, 8.30 mmol) was added to thereaction mixture. The progress of the reaction was followed by HPLCanalysis. After 2 h at ambient temperature, the reaction was quenchedwith water (5 mL) and then concentrated to half its volume under vacuum.The reaction mixture was diluted with dichloromethane (15 mL) and washedwith 1N sodium hydroxide (2×10 mL) and then brine (5 mL). The organiclayer was dried over magnesium sulfate and concentrated to yield thetitle compound.

Preparation 47

2-Benzyloxy-5-(2-bromoacetyl)benzoic Acid Methyl Ester (a)2-Benzyloxy-5-acetylbenzoic Acid Methyl Ester

Methyl 5-acetylsalicylate (100 g, 0.515 mol) was dissolved inacetonitrile (1 L) in a 2 L flask under reflux conditions and a nitrogenatmosphere. Potassium carbonate (213.5 g, 1.545 mol) was addedportion-wise over 15 min. Benzyl bromide (67.4 mL, 0.566 mol) was addedusing a dropping funnel over 15 min. The reaction was heated to 85° C.for 9 h, and then filtered and rinsed with acetonitrile (100 mL). Thesolution was concentrated to about 300 mL volume under reduced pressureand partitioned between water (1 L) and ethyl acetate (1 L). The organiclayer was washed with saturated sodium chloride (250 mL), dried usingmagnesium sulfate (75 g), and then filtered and rinsed with ethylacetate (100 mL). The organic layer was concentrated to give2-benzyloxy-5-acetylbenzoic acid methyl ester as a solid (100% yield).

(b) 2-Benzyloxy-5-(2-bromoacetyl)benzoic Acid Methyl Ester

The product of step (a) (10.0 g, 35.2 mmol) was dissolved in chloroform(250 mL) in a 500 mL flask under a nitrogen atmosphere. Bromine (1.63mL, 31.7 mmol) dissolved in chloroform (50 mL) was added using adropping funnel over 30 min. The reaction mixture was stirred for 2.5 hand then concentrated to give a solid. The solid was dissolved intoluene (150 mL) with some gentle heat, followed by the addition ofethyl ether (150 mL) to yield the title compound as a crystalline solid(55% yield).

Preparation 48

5-[2-(Benzyl-{9-[4-(3-biphenyl-2-ylureido)piperidin-1-yl]nonyl}amino)acetyl]-2-benzyloxybenzoicAcid Methyl Ester

The product of Preparation 47 (371 mg, 1.00 mmol) was added to asolution of the product of Preparation 46 (448 mg, 0.85 mmol) indimethyl sulfoxide (4.5 mL) followed by the addition of potassiumcarbonate (234 mg, 1.7 mmol). The reaction mixture was stirred at 40° C.for 6 h, at which time the product of Preparation 46 was no longerobserved by HPLC analysis. The reaction mixture was cooled to ambienttemperature and filtered, and then diluted with ethanol (4 mL). Sodiumborohydride (63 mg, 1.7 mmol) was added to the reaction mixture and thereaction was stirred at ambient temperature for 24 h. The reactionmixture was quenched with 0.5 M ammonium chloride (5 mL) and extractedinto ethyl acetate (2×10 mL). The combined organic layers were washedwith saturated sodium bicarbonate (10 mL) and then with brine (5 mL).The organic layer was dried over magnesium sulfate and the solvent wasremoved under reduced pressure. The crude residue was purified bychromatography on silica gel (3% methanol in chloroform) to give thetitle compound.

Preparation 49

1-[1-(9-{Benzyl-[2-(4-benzyloxy-3-hydroxymethylphenyl)-2-hydroxyethyl]amino}nonyl)piperidin-4-yl]-3-biphenyl-2-ylurea

A solution of the product of Preparation 48 (163 mg, 0.20 mmol) intetrahydrofuran (1.00 mL) was cooled to 0° C. Lithium aluminium hydride(1.0 M in THF; 0.50 mL, 0.50 mmol) was added dropwise to the mixture.After 1 h, the reaction mixture was quenched with water (1 mL) anddiluted with ethyl acetate (2 mL). The organic layer was washed withbrine, dried over magnesium sulfate, and the organic extracts werecombined and concentrated to give the title compound.

Example 131-Biphenyl-2-yl-3-(1-{9-[2-hydroxy-2-(4-hydroxy-3-hydroxymethylphenyl)ethylamino]nonyl}piperidin-4-yl)ureaDihydrochloride

A solution of the product of Preparation 49 (130 mg, 0.16 mmol) inisopropanol (0.80 ml) was flushed with nitrogen for ten minutes and thenpalladium (10 wt. % (dry basis) on activated carbon (60 mg) was added.The reaction flask was purged with nitrogen and then a balloon filledwith hydrogen was attached to the flask and the reaction mixture wasstirred under an atmosphere of hydrogen. After 72 h, the reactionmixture was filtered and concentrated and the residue was purified bypreparative HPLC. The resulting ditrifluoroacetic acid salt of the titlecompound was dissolved in 1N hydrochloric acid (5 mL) and lyophilized toyield the title compound as its dihydrochloride salt.

Preparation 50

5-[(R)-2-[(3-Aminomethylcyclohexylmethyl)amino]-1-(tert-butyldimethylsilanyloxy)ethyl]-8-benzyloxy-1H-quinolin-2-one

A stirred solution of the product of Preparation 13 (1.46 g, 3 mmol) and1,3-cyclohexanebis(methylamine) (426 mg, 3 mmol) in DMSO (3 mL) washeated at 100° C. for 6 h. The reaction mixture was allowed to cool andit was then diluted with dichloromethane (20 mL) and washed withsaturated aqueous sodium bicarbonate solution (10 mL). The organic layerwas dried (MgSO₄) and the solvent was removed under reduced pressure.The crude residue was purified by flash chromatography (10% MeOH/DCM and0.5% NH₄OH) to give the title compound as a solid (775 mg, 50% yield).MS m/z: [M+H⁺] calcd for C₃₂H₄₇N₃O₃Si 550.3. found 550.6.

Preparation 51

Biphenyl-2-ylcarbamic Acid1-{2-[(3-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydro-quinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]methyl}-cyclohexylmethyl)carbamoyl]ethyl}piperidin-4-ylEster

To a stirred solution of the product of Preparation 50 (552 mg, 1.01mmol), the product of Preparation 20 (309 mg, 0.84 mmol) and HATU (384mg, 1.01 mmol) in DMF (1.68 mL) was added DIPEA (190 μL, 1.09 mmol). Thereaction mixture was stirred at 50° C. overnight and then the solventwas removed under reduced pressure. The resulting residue was dissolvedin dichloromethane (20 mL) and washed with saturated aqueous sodiumbicarbonate solution (10 mL). The organic phase was dried (magnesiumsulfate) and the solvent was removed under reduced pressure. The crudeproduct was purified by flash chromatography (5-10% MeOH/DCM) to givethe title compound as a solid (267 mg, 36% yield). LCMS (10-70)R_(t)=5.04. MS m/z: [M+H⁺] calcd for C₅₃H₆₉N₅O₆Si 900.5. found 900.6.

Preparation 52

Biphenyl-2-ylcarbamic Acid1-{2-[(3-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]methyl}cyclohexylmethyl)carbamoyl]ethyl}piperidin-4-ylEster

To a stirred solution of the product of Preparation 51 (267 mg, 0.30mmol) in dichloromethane (3 mL) was added triethylamine trihydrofluoride(98 μL, 0.6 mmol). The reaction mixture was stirred for 10 h and then itwas diluted with dichloromethane (10 mL) and washed with saturatedaqueous sodium bicarbonate solution (5 mL). The organic phase was dried(magensium sulfate) and the solvent was removed under reduced pressureto give the title compound as a solid (236 mg, 100% yield). MS m/z:[M+H⁺] calcd for C₄₇H₅₅N₅O₆ 786.4. found 786.5.

Example 14 Biphenyl-2-ylcarbamic Acid1-{2-[(3-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-cyclohexylmethyl)carbamoyl]ethyl}-piperidin-4-ylEster

Palladium (10 wt. % (dry basis) on activated carbon) (120 mg) was addedto a stirred solution of the product of Preparation 52 (236 mg, 0.30mmol) in ethanol (3 mL). The reaction mixture was placed under ahydrogen atmosphere and stirred overnight. The reaction mixture was thenfiltered and the solvent was removed under reduced pressure. The cruderesidue was purified by preparative HPLC to give the title compound (27mg, 2 TFA salt). HPLC (10-70) R_(t)=2.76. MS m/z: [M+H⁺] calcd forC₄₀H₄₉N₅O₆ 696.4. found 696.6.

Preparation 53

Biphenyl-2-ylcarbamic Acid1-{2-[((1R,3S)-3-Aminocyclopentanecarbonyl)amino]-ethyl}piperidin-4-ylEster

To a stirred solution of the product of Preparation 39 (318 mg, 0.94mmol), (1R,3S)-3-tert-butoxycarbonylaminocyclopentanecarboxylic acid(258 mg, 1.1 mmol) and HATU (428 mg, 1.1 mmol) in DMF (5 mL) was addedDIPEA (245 μL, 1.09 mmol). The reaction mixture was stirred at roomtemperature overnight and then the solvent was removed under reducedpressure. The resulting residue was dissolved in dichloromethane (20 mL)and washed with saturated aqueous sodium bicarbonate solution (10 mL).The organic layer was dried (magnesium sulfate) and the solvent wasremoved under reduced pressure. The crude product was purified by flashchromatography (5-10% MeOH/DCM) and then dissolved in a trifluoroaceticacid/DCM mixture (1 mL/5 mL) and stirred at room temperature for 1 h.The solvent was removed under reduced pressure. The residue wasdissolved in dichloromethane (20 mL) and washed with 1M sodium hydroxide(10 mL), dried (magnesium sulfate) and the solvent reduced to yield thetitle compound (167 mg, 39% yield).

Preparation 54

Biphenyl-2-ylcarbamic Acid1-[2-({(1R,3S)-3-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]-cyclopentanecarbonyl}amino)ethyl]piperidin-4-ylEster

A stirred solution of the product of Preparation 53 (167 mg, 0.38 mmol)and the product of Preparation 13 (92 mg, 0.19 mmol) in DMSO (0.38 mL)was heated at 90° C. for 5 h. The solution was cooled and diluted withethyl acetate (10 mL) and then washed with saturated aqueous sodiumbicarbonate (5 mL). The organic phase was dried (magnesium sulfate) andthe solvent was removed under reduced pressure. The crude product waspurified by flash chromatography (5-10% MeOH/DCM) to yield the titlecompound (343 mg, 100% yield). LCMS (10-70) R_(t)=4.97. MS m/z: [M+H⁺]calcd for C₅₀H₆₃N₅O₆Si 858.5. found 858.8.

Preparation 55

Biphenyl-2-ylcarbamic Acid1-[2-({(1R,3S)-3-[(R)-2-(8-benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]cyclopentanecarbonyl}amino)ethyl]-piperidin-4-ylEster

To a stirred solution of the product of Preparation 54 (343 mg, 0.4mmol) in THF (2 mL) was added triethylamine trihydrofluoride (130 μL,0.8 mmol). The reaction mixture was stirred for 10 h and was thendiluted with EtOAc (10 mL). The reaction mixture was washed withsaturated aqueous sodium bicarbonate solution (5 mL) and then theorganic phase was dried (magnesium sulfate) and the solvent was removedunder reduced pressure to give the title compound as a solid (298 mg,100% yield). HPLC (10-70) R_(t)=2.8. MS m/z: [M+H⁺] calcd for C₄₄H₄₉N₅O₆744.4. found 744.4.

Example 15 Biphenyl-2-ylcarbamic Acid1-[2-({(1R,3S)-3-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)ethylamino]-cyclopentanecarbonyl}amino)ethyl]-piperidin-4-ylEster Ditrifluoroacetate

To a stirred solution of the product of Preparation 55 (236 mg, 0.40mmol) in ethanol (3 mL) was added palladium (10 wt. % (dry basis) onactivated carbon (120 mg). The reaction mixture was placed under ahydrogen atmosphere and stirred overnight. The reaction mixture wasfiltered and the solvent was removed under reduced pressure. The cruderesidue was purified by preparative HPLC to give the title compound (3mg, 2 TFA salt). HPLC (5-75) R_(t)=2.18. MS m/z: [M+H⁺] calcd forC₃₇H₄₅N₅O₆ 656.3. found 656.2.

Preparation 56

4-(tert-Butoxycarbonylaminomethyl)-2-chlorophenylamine

A stirred solution of 4-aminomethyl-2-chlorophenylamine (940 mg, 6 mmol)and di-tert-butyl dicarbonate (1.44 g, 6.6 mmol) in dichloromethane (30mL) was stirred at room temperature for 4 h, at which time the reactionwas determined to be complete by LCMS. The reaction mixture was thenwashed with saturated aqueous sodium bicarbonate (15 mL) and the organiclayer was dried over sodium sulfate and the solvent was removed underreduced pressure. The resulting orange solid was recrystallized fromethyl acetate to give the title intermediate as a white solid (˜100%yield).

Preparation 57

N-[4-(tert-Butoxycarbonylaminomethyl)-2-chlorophenyl]acrylamide

To a stirred solution of the product of Preparation 56 (1.54 g, 6.0mmol) in a mixture of diethyl ether (35 mL) and 1 M sodium hydroxide (35mL) was added dropwise acryloyl chloride (687 μL, 8.45 mmol). After 1 h,the organic layer was separated, dried (Na₂SO₄) and the solvent wasremoved under reduced pressure to give the title intermediate as a whitesolid (1.8 g, 96% yield).

Preparation 58

Biphenyl-2-ylcarbamic Acid1-[2-(4-(tert-Butoxycarbonylaminomethyl)-2-chlorophenylcarbamoyl)ethyl]piperidin-4-ylEster

A solution of the product of Preparation 8 (1.04 g, 3.5 mmol) and theproduct of Preparation 57 (1.19 g, 3.85 mmol) in a mixture ofdichloromethane and methanol (12 mL, 1:1) was heated at 60° C. for 12 h.The reaction mixture was allowed to cool and the solvent was removedunder reduced pressure. The crude material was purified by columnchromatography (5-10% MeOH/DCM) to give the title intermediate as awhite solid (2.00 g, 94% yield).

Preparation 59

Biphenyl-2-ylcarbamic Acid1-[2-(4-Aminomethyl-2-chlorophenylcarbamoyl)ethyl]-piperidin-4-yl Ester

A solution of the product of Preparation 58 (2.00 g, 3.3 mmol) wasstirred in dichloromethane (24 mL) and TFA (8 mL) for 1 h and then thesolvent was removed under reduced pressure. The crude reaction mixturewas dissolved in dichloromethane (30 mL) and washed with 1 M sodiumhydroxide (2×30 mL). The organic layer was dried (Na₂SO₄) and thesolvent was removed under reduced pressure to give the titleintermediate as an oily white solid (1.46 g, 88% yield).

Preparation 60

Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]methyl}-2-chlorophenylcarbamoyl)-ethyl]piperidin-4-ylEster

A stirred solution of the product of Preparation 59 (1.41 g, 2.79 mmol)and the product of Preparation 13 (680 mg, 1.39 mmol) in DMSO (1.39 mL)was heated at 90° C. for 8 h and then cooled to room temperature. Thereaction mixture was diluted with ethyl acetate/chloroform (20 mL, 1/1)and the organic layer was washed with saturated aqueous sodiumbicarbonate (10 mL), dried (Na₂SO₄) and the solvent removed underreduced pressure. The resulting crude residue was purified by columnchromatography (5-10% MeOH/DCM) to give the title intermediate as awhite solid (1.12 g, 88% yield). MS m/z M+H⁺=914.9.

Preparation 61

Biphenyl-2-yl-Carbamic Acid1-[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]methyl}-2-chloro-phenylcarbamoyl)ethyl]piperidin-4-ylEster

To a stirred solution of the product of Preparation 60 (1.12 g, 1.23mmol) in dichloromethane (12 mL) was added Et₃N.3HF (401 μL, 0.6 mmol).The reaction mixture was stirred for 10 h and then diluted withdichloromethane (10 mL). This mixture was washed with saturated aqueoussodium bicarbonate solution (5 mL) and the organic layer dried (Na₂SO₄)and the solvent removed under reduced pressure to give the titleintermediate as a white solid (959 mg, 100% yield). MS m/z M+H⁺=800.5.

Example 16 Biphenyl-2-ylcarbamic Acid1-[2-(2-Chloro-4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-ylEster Ditrifluoroacetate

To a stirred solution of the product of Preparation 61 (959 mg, 1.2mmol) in ethanol (12 mL) was added Pd/C (290 mg) and the reactionmixture was placed under a hydrogen atmosphere and stirred overnight.The reaction mixture was then filtered and the solvent removed underreduced pressure. The crude residue was purified by preparative HPLC togive the title compound (67 mg, 2 TFA salt). HPLC (10-70) R_(t)=2.76; MSm/z M+H⁺=710.6.

Preparation 62

2-Chloroethanesulfonic Acid (5-tert-Butoxycarbonylaminopentyl)amide

To a stirred solution of 5-(tert-butoxycarbonylamino)pentylamine (1.00g, 4.94 mmol) and triethylamine (689 μg, 4.94 mmol) in dichloromethane(22 mL) at 0° C. was added 2-chloro-1-ethanesulfonyl chloride (470 μL,4.50 mmol). The reaction mixture was stirred for 2 h at room temperatureand then washed with saturated aqueous sodium bicarbonate solution (15mL). The organic layer was dried (Na₂SO₄) and the solvent was removedunder reduced pressure to give the title compound (100% yield), whichwas used in the next step without further purification.

Preparation 63

Biphenyl-2-ylcarbamic Acid1-[2-(5-tert-Butoxycarbonylaminopentylsulfamoyl)-ethyl]piperidin-4-ylEster

A solution of the product of Preparation 8 (1.33 g, 3.5 mmol) and theproduct of Preparation 62 (1.62 g, 4.94 mmol) in dichloromethane andmethanol (22 mL, 1:1) was heated at 60° C. for 5 h. The reaction mixturewas allowed to cool to room temperature and the solvent was removedunder reduced pressure. The crude residue was dissolved indichloromethane (20 mL) and washed with saturated aqueous sodiumbicarbonate solution (10 mL). The organic layer was then dried (Na₂SO₄)and solvent removed under reduced pressure. The crude residue waspurified by column chromatography (5-10% MeOH/DCM) to give the titleintermediate as a white solid (1.6 g, 55%). MS m/z M+H⁺=589.6.

Preparation 64

Biphenyl-2-ylcarbamic Acid1-[2-(5-Aminopentylsulfamoyl)ethyl]piperidin-4-yl Ester

A solution of the product of Preparation 63 (1.6 g, 2.72 mmol) wasstirred in dichloromethane (21 mL) and TFA (7 mL) for 1 h and then thesolvent was removed under reduced pressure. The crude reaction mixturewas dissolved in dichloromethane (30 mL) and washed with 1 M sodiumhydroxide (2×30 mL). The organic layer was dried (Na₂SO₄) and thesolvent was then removed under reduced pressure to give the titleintermediate as an oily white solid (1.19 g, 90% yield).

Preparation 65

Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]pentylsulfamoyl}ethyl)piperidin-4-ylEster

A stirred solution of the product of Preparation 64 (917 mg, 1.88 mmol)and the product of Preparation 13 (460 mg, 0.94 mmol) in DMSO (0.92 mL)was heated at 90° C. for 8 h and then cooled to room temperature. Thereaction mixture was diluted with ethyl acetate/chloroform (20 mL, 1/1)and the organic layer was washed with saturated aqueous sodiumbicarbonate solution (10 mL), dried (Na₂SO₄) and the solvent was removedunder reduced pressure. The resulting crude residue was purified bycolumn chromatography (3-6% MeOH/DCM) to give the title intermediate asa white solid (500 mg, 60% yield). MS m/z M+H⁺=896.9.

Preparation 66

Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]pentylsulfamoyl}ethyl)piperidin-4-ylEster

To a stirred solution of the product of Preparation 65 (500 mg, 0.56mmol) in dichloromethane (5.6 mL) was added triethylaminetrihydrofluoride (183 μL, 1.12 mmol). The reaction mixture was stirredfor 10 h and dichloromethane (10 mL) was added. The resulting mixturewas washed with saturated aqueous sodium bicarbonate solution (5 mL).The organic layer was dried (Na₂SO₄) and the solvent was removed underreduced pressure to give the title intermediate as a yellow solid (437mg, 100% yield. MS m/z M+H⁺=782.8.

Example 17 Biphenyl-2-ylcarbamic Acid1-(2-{5-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylsulfamoyl}ethyl)piperidin-4-ylEster Ditrifluoroacetate

To a stirred solution of the product of Preparation 66 (437 mg, 0.56mmol) in ethanol/methanol (5.6 mL, 1/1) was added Pd/C (131 mg) and thereaction mixture placed under a hydrogen atmosphere and stirredovernight. The reaction mixture was then filtered and the solvent wasremoved under reduced pressure. The crude residue was purified bypreparative HPLC to give the title compound as the ditrifluoroaceticacid salt (71 mg). HPLC (10-70) R_(t)=2.59; MS m/z M+H⁺=692.6.

Preparation 67

Biphenyl-2-ylcarbamic Acid1-{2-[(4-Formylbenzenesulfonyl)methylamino]-ethyl}piperidin-4-yl Ester

To a stirred solution of the product of Preparation 26 (350 mg, 1 mmol)and triethylamine (167 μL, 1.2 mmol) in dichloromethane (5 mL) was added4-formylbenzenesulfonyl chloride (225 mg, 1.1 mmol). After 1 h at roomtemperature, the reaction was complete by MS and the reaction mixturewas then washed with saturated aqueous sodium bicarbonate solution (5mL). The organic layer was then dried (Na₂SO₄) and solvent removed underreduced pressure to give the title intermediate (323 mg, 62% yield). MSm/z M+H⁺=522.4.

Preparation 68

Biphenyl-2-ylcarbamic Acid1-{2-[(4-{[(R)-2-(tert-Butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzenesulfonyl)-methylamino]ethyl}piperidin-4-ylEster

A solution of5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-one(293 mg, 0.74 mmol) and the product of Preparation 67 in dichloromethaneand methanol (6.2 mL, 1/1) was stirred at room temperature for 1 h andthen sodium triacetoxyborohydride (394 mg, 1.86 mmol) was added. Thereaction mixture was stirred for 4 h at which time the reaction wasdetermined to be complete by MS. The reaction mixture was then acidifiedwith concentrated hydrochloric acid and the solvent was removed underreduced pressure to provide the title compound, which was used in thenext step without further purification. MS m/z M+H⁺=840.8.

Example 18 Biphenyl-2-ylcarbamic Acid1-{2-[(4-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzenesulfonyl)methylamino]ethyl}-piperidin-4-ylEster Ditrifluoroacetate

A stirred solution of the product of Preparation 68 (520 mg, 0.62 mmol)in 1M hydrochloric acid (5 mL) and acetonitrile (5 mL) was heated at 60°C. for 8 h. The reaction mixture was cooled to room temperature and thesolvent was removed under reduced pressure. The crude residue waspurified by preparative HPLC to give the title compound as theditrifluoroacetic acid salt (220 mg). HPLC (10-70) R_(t)=2.77; MS m/zM+H⁺=726.7.

Preparation 69

(3-Aminomethylphenyl)methanol Hydrochloride (a)(3-tert-Butoxycarbonylmethylphenyl)methanol

Borane dimethyl sulfide (2.05 mL, 21.6 mmol) was added to a solution of3-(tert-butoxycarbonylaminomethyl)benzoic acid (1.81 g, 7.20 mmol) intetrahydrofuran (24 mL). and the resulting mixture was stirred at roomtemperature for 3 hours. The reaction mixture was then diluted withethyl acetate (20 mL) and the layers were separated. The organic layerwas washed with saturated sodium bicarbonate, saturated sodium chloride,dried over magnesium sulfate and concentrated to give the title compoundas a yellow oil (1.71 g).

(b) (3-Aminomethylphenyl)methanol Hydrochloride

To the product of step (a) (1.71 g, 7.2 mmol) was added a solution of 4M hydrochloric acid in dioxane (9 mL, 36 mmol) and the resulting mixturewas stirred at room temperature for 1 h. The reaction mixture was thenconcentrated and the residue was diluted with diethyl ether (50 mL) andfiltered to provide the title compound as a white solid (1.09 g).

Preparation 70

Biphenyl-2-ylcarbamic Acid1-{2-[3-(3-Hydroxymethylbenzyl)ureido]ethyl}piperidin-4-yl Ester

A 0.2 M solution of the product of Preparation 35 (760 mg, 2.24 mmol) inN,N-dimethylformamide was added dropwise to a solution of1,1′-carbonyldiimidazole (364 mg, 2.24 mmol) and diisopropylethylamine(0.31 mL, 2.24 mmol) in N,N-dimethylformamide (11 mL) and the resultingmixture was stirred at room temperature for 2 h. Diisopropylethylamine(0.31 mL, 2.24 mmol) and the product of Preparation 69 (578 mg, 3.4mmol) were added and this mixture was stirred at 50° C. for 12 hours.The reaction mixture was then concentrated to dryness and the residuewas diluted with dichloromethane (20 mL) and this solution was washedwith saturated sodium bicarbonate (2×), saturated sodium chloride, driedover magnesium sulfate, and concentrated to provide the title compound(1.12 g). LCMS (2-90) R_(t)=4.01 min.; MS m/z M+H=503.5.

Preparation 71

Biphenyl-2-ylcarbamic Acid1-{2-[3-(3-Formylbenzyl)ureido]ethyl}piperidin-4-yl Ester

A solution of the product of Preparation 70 (1.12 g, 2.23 mmol) indichloromethane (11.1 mL) was cooled to 0° C. and diisopropylethylamine(1.17 mL, 6.70 mmol) and dimethyl sulfoxide (0.949 mL, 13.4 mmol) wereadded. After about 10 minutes, pyridine sulfur trioxide complex (1.06 g,6.70 mmol) was added and the resulting mixture was stirred at 0° C. for2 h. The reaction was then quenched with water (15 mL) and the organiclayer was washed with cold water (3×), dried over magnesium sulfate andconcentrated to provide the title compound as a yellow crisp (609 mg).LCMS (2-90) R_(t)=4.13 min; MS m/z M+H=501.3.

Preparation 72

Biphenyl-2-ylcarbamic Acid1-{2-[3-(3-{[(R)-2-(tert-butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzyl)ureido]ethyl}-piperidin-4-ylEster

5-[(R)-2-Amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-one(575 mg, 1.40 mmol) was added to a solution of the product ofPreparation 71 (609 mg, 1.2 mmol) and diisopropylamine (0.25 mL, 1.40mmol) in dichloromethane (6 mL) and the resulting mixture was stirred atroom temperature for 45 min. Sodium triactetoxyborohydride (385 mg, 1.80mmol) was then added and this mixture was stirred at room temperaturefor 12 h. The reaction was then quenched with 10% aqueous hydrochloricacid (5 mL) and the layers were separated. The organic layer was washedwith saturated sodium bicarbonate, saturated sodium chloride, dried overmagnesium sulfate and concentrated to give the title compound (1.1 g).HPLC (10-70) R_(t)=3.55 min; MS m/z M+H=819.7.

Example 19 Biphenyl-2-ylcarbamic Acid1-{2-[3-(3-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzyl)ureido]ethyl}piperidin-4-ylEster Ditrifluoroacetate

Triethylamine trihydrofluoride (2.4 mL, 13.6 mmol) was added to asolution of the product of Preparation 72 (1.1 g, 1.36 mmol) indichloromethane (2 mL) and the resulting mixture was stirred at roomtemperature for 15 h. The reaction mixture was then concentrated undervacuum to dryness and the residue was dissolved in a 1:1 mixture ofwater and acetonitrile with 0.1% TFA and this mixture was purified byHPLC (5-35 over 60 min) to provide the title compound as theditrifluoroacetate salt (296 mg, 99% purity). MS m/z M+H=705.6.

Preparation 73

Biphenyl-2-ylcarbamic Acid 1-[(E)-3-(4-Nitrophenyl)allyl]piperidin-4-ylEster

The product of Preparation 8 (2.96 g, 0.01 mol) andp-nitrocinnamaldehyde (1.77 g, 0.01 mol) were stirred in 50 mL ofdichloromethane for 2 h. Sodium triacetoxyborohydride (6.33 g, 0.03 mol)was added and the resulting mixture was stirred for 2 h. The reactionwas then quenched with 10 mL of water and this mixture was diluted withdichloromethane (100 mL). The organic layer was washed with saturatedsodium bicarbonate (2×), brine, dried over Na₂SO₄, filtered andconcentrated to provide the title compound as a yellow foam (3.8 g, 80%yield).

Preparation 74

Biphenyl-2-ylcarbamic Acid 1-[3-(4-Aminophenyl)propyl]piperidin-4-ylEster

The product of Preparation 73 (2.5 g, 5.4 mmol) was dissolved in 100 mLof ethanol and the resulting solution was purged with nitrogen for 30min. Palladium on carbon (2.5 g; 50% w/w water; 10% Pd; 1.1 mmol Pd) wasthen added while degassing with nitrogen. This mixture was then placedunder hydrogen (50 psi) until hydrogen was no longer consumed (˜30minutes). The mixture was then purged with nitrogen, filtered throughCelite and concentrated. The residue was dissolved in ethyl acetate andthis mixture was washed with saturated sodium bicarbonate (2×), brine,dried (Na₂SO₄), filtered and concentrated to provide the title compound(2.08 g, 90% yield). MS m/z M+H=430.5.

Preparation 75

Biphenyl-2-ylcarbamic Acid1-{3-[4-(4-{2-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydro-quinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]ethyl}phenylamino)-phenyl]propyl}piperidin-4-ylEster

To a 25 mL round-bottomed flask was added the product of Preparation 74(400 mg, 0.8 mmol);8-benzyloxy-5-[(R)-2-[2-(4-bromophenyl)ethylamino]-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one(769 mg, 1.2 mmol); tris(dibenzylideneacetone)dipalladium(0) (73 mg,0.08 mmol, 20% Pd); and 2-(dicyclohexylphosphino)biphenyl (84 mg, 0.24mmol). This mixture was purged with nitrogen and then dry, degassedtoluene (8 mL, 0.1 M) was added and the resulting mixture was heated at70° C. for 30 min. Sodium tert-butoxide (382 mg, 4.0 mmol) was thenadded, and the temperature was raised to 95° C. for 4 h, at which timeLCMS showed complete consumption of the product of Preparation 74 and alarge product peak (M+H=956.7). The reaction mixture was then cooled toroom temperature and diluted with ethyl acetate. This mixture was washedwith saturated sodium bicarbonate (2×), brine, dried (Na₂SO₄), filteredand concentrated to provide the title compound (1.5 g), which was usedwithout further purification.

Preparation 76

Biphenyl-2-ylcarbamic Acid1-{3-[4-(4-{2-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydro-quinolin-5-yl)-2-hydroxyethylamino]ethyl}phenylamino)phenyl]propyl}piperidin-4-ylEster

The product of Preparation 75 was dissolved in dichloromethane (10 mL)and triethylamine trihydrofluoride (10 eq.) was added. The reactionmixture was stirred overnight and then diluted with dichloromethane andthe organic layer was washed with saturated sodium bicarbonate (2×),brine, dried (Na₂SO₄), filtered and concentrated to provide 1.3 g ofcrude product. This material was purified by silica gel chromatography(DCM, incrementally to 50% methanol) to provide the title compound (300mg, about 75% purity), which was used without further purification.

Example 20 Biphenyl-2-ylcarbamic acid1-{3-[4-(4-{2-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)phenyl]propyl}piperidin-4-ylester Ditrifluoroacetate

The product of Preparation 76 (300 mg) was dissolved in 10 mL of ethanoland this mixture was purged with nitrogen for 15 minutes. Palladium oncarbon (10% Pd, 50% w/w water, 0.2 eq. Pd) was added while degassing.The resulting mixture was then placed under 1 atm. of hydrogen for 2 h,at which time the reaction was complete by LCMS. The solution was thenpurged with nitrogen for 15 min and then filtered through Celite andconcentrated. The resulting residue was purified by prep HPLC to affordthe title compound as the ditrifluoroacetate salt (59 mg, >95% purity).MS m/z M+H=752.8.

Preparation 77

Biphenyl-2-ylcarbamic Acid1-[2-Fluoro-3-(4-hydroxymethylpiperidin-1-ylmethyl)-benzyl]piperidin-4-ylEster

The product of Preparation 8 (500 mg, 1.69 mmol),2,6-bis(bromomethyl)-1-fluorobenzene (476 mg, 1.69 mmol,piperidin-4-ylmethanol (195 mg, 1.69 mmol) and potassium carbonate (466mg, 3.37 mmol) were suspended in acetonitrile (5 mL) and stirred at roomtemperature for 18 h. The reaction mixture was then concentrated and theresidue was dissolved in dichloromethane/water. The layers wereseparated and the organic layer was washed with water (2×), brine, dried(MgSO₄) and concentrated. The crude material was purified by silica gelcolumn chromatography eluting with 3% methanol/chloroform to give thetitle compound as a white foam (282 mg). MS m/z M+H=532.3.

Preparation 78

Biphenyl-2-ylcarbamic Acid1-[2-Fluoro-3-(4-formylpiperidin-1-ylmethyl)benzyl]-piperidin-4-yl Ester

The product of Preparation 77 (282 mg, 0.53 mmol) was dissolved indichloromethane and to this mixture was added diisopropylethylamine (280μL, 1.6 mmol) and dimethyl sulfoxide (115 μL, 1.6 mmol). The reactionmixture was cooled to −15° C. under nitrogen and pyridine sulfurtrioxide complex (255 mg, 1.6 mmol) was added and the resulting mixturewas stirred for 40 min. The reaction was then quenched with water andthe layers were separated. The organic layer was washed with aqueousNaH₂PO₄ (1M×3), brine, dried (MgSO₄) and concentrated to provide thetitle compound as a foam (253 mg). MS m/z M+H=530.4.

Example 21 Biphenyl-2-ylcarbamic Acid1-[2-Fluoro-3-(4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}piperidin-1-ylmethyl)benzyl]piperidin-4-ylEster Ditrifluoroacetate

The product of Preparation 78 (253 mg, 0.48 mmol) was dissolved in a 1:1mixture of dichloromethane and methanol (6 mL) and to this mixture wasadded5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-oneacetate (228 mg, 0.58 mmol) and sodium triacetoxyborohydride (317 mg,1.5 mmol). The reaction mixture was stirred under nitrogen at roomtemperature for 18 h and then concentrated. The residue was dissolved ina 2:3 mixture of acetonitrile and aqueous 6 N hydrochloric acid, andthis mixture was heated at 55° C. for 4 hours. The reaction mixture wasthen concentrated and the residue was dissolved inwater/acetonitrile/trifluoroacetic acid (1:1:0.005) and purified byreverse phase column chromatography to afford the title compound as awhite solid (175 mg). MS m/z M+H=734.5.

Preparation 79

2-[4-(3-Bromopropoxy)phenyl]ethanol

To a solution of 4-hydroxyphenethyl alcohol (4.37 g, 31.0 mmol) andpotassium carbonate (6.55 g, 47.0 mmol) in acetonitrile (62.0 mL) wasadded 1,3 dibromopropane (31.0 mL, 316 mmol). The reaction mixture washeated to 70° C. for 12 hours and then cooled to room temperature,filtered and concentrated under vacuum. The resulting oil was purifiedby silica gel chromatography using a mixture of 4:1 hexanes and ethylacetate to give the title compound (6.21 g) as a white solid.

Preparation 80

Biphenyl-2-ylcarbamic Acid1-{3-[4-(2-Hydroxyethyl)phenoxy]propyl}piperidin-4-yl Ester

To a solution of the product of Preparation 79 (1.11 g, 4.30 mmol) anddiisopropylethylamine (0.90 mL, 5.10 mmol) in acetonitrile (21.5 mL) wasadded the product of Preparation 8 (1.27 g, 4.30 mmol) and the resultingmixture was stirred at 60° C. for 12 h. The reaction mixture was thendiluted with dichloromethane (20 mL) and washed with saturated sodiumbicarbonate (25 mL), saturated sodium chloride (25 mL), dried overmagnesium sulfate and concentrated to provide the title compound (1.98g, 85% purity). MS m/z M+H=475.5.

Preparation 81

Biphenyl-2-ylcarbamic Acid1-{3-[4-(2-Oxoethyl)phenoxy]propyl}piperidin-4-yl Ester

A solution of the product of Preparation 80 (723 mg, 1.53 mmol) anddichloromethane (75 mL) was cooled to about 5° C. anddiisopropylethylamine (798 mL, 4.58 mmol) and dimethyl sulfoxide (649mL, 9.15 mmol) were added. Pyridine sulfur trioxide (728 mg, 4.58 mmol)was then added and the resulting mixture was stirred at 5° C. for 45min. The reaction mixture was then diluted with dichloromethane (20 mL)and washed with saturated sodium bicarbonate (25 mL), saturated sodiumchloride (25 mL), dried over magnesium sulfate and concentrated toprovide the title compound (604 mg). MS m/z M+H=473.4.

Preparation 82

Biphenyl-2-ylcarbamic Acid1-[3-(4-{2-[(R)-2-(tert-butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}-phenoxy)propyl]piperidin-4-ylEster

The product of Preparation 81 (604 mg, 1.28 mmol) was dissolved inmethanol (6.4 mL) and5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-one(605 mg, 1.53 mmol) and diisopropylethylamine (0.27 mL, 1.53 mmol) wereadded. Sodium triacetoxyborohydride (405 mg, 1.91 mmol) was then addedand the reaction mixture was stirred at room temperature for 3 h. Thereaction mixture was then concentrated to dryness and the residue wasdiluted with ethyl acetate (20 mL) and this solution was washed withsaturated sodium bicarbonate (25 mL), saturated sodium chloride (25 mL),dried over magnesium sulfate and concentrated to give the title compound(704 mg). MS m/z M+H=791.8.

Example 22 Biphenyl-2-ylcarbamic Acid1-[3-(4-{2-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenoxy)propyl]piperidin-4-ylEster Ditrifluoroacetate

Triethylamine trihydrofluoride (1.5 mL, 8.87 mmol) was added to asolution of the product of Preparation 82 (702 mg, 0.89 mmol) indichloromethane (4.5 mL) and the resulting mixture was stirred at roomtemperature for 24 h. The mixture was then concentrated under vacuum andpurified by HPLC (2-35 over 90 min) to give the title compound (92 mg)as a white powder. MS m/z M+H=677.4.

Preparation 83

Methyl 4-Iodophenylacetate

To a stirred solution of 4-iodophenylacetic acid (5.0 g, 19.1 mmol) inMeOH (200 mL) was added 4N hydrochloric acid in dioxane (10 mL). Thereaction mixture was stirred for 24 h at room temperature and then thesolvent was removed under reduced pressure to give the title compound(5.17 g, 98% yield), which was used without further purification.

Preparation 84

Methyl [4-(4-Hydroxybut-1-ynyl)phenyl]acetate

To a stirred solution of the product of Preparation 83 (4.5 g, 16.3mmol) in diethylamine (100 mL) was added but-3-yn-1-ol (1.9 mL, 32.6mmol), Pd(PPh₃)₂Cl₂ (500 mg, 1.63 mmol) and CuI (154 mg, 0.815 mmol) andresulting mixture was stirred for 17 h at room temperature. The solventwas then removed under reduced pressure and the residue was dissolved indiethyl ether (200 mL) and this solution was filtered to remove salts.The solvent was then removed under reduced pressure and the crudeproduct was purified by silica gel chromatography (60% EtOAc/Hexane) toafford the title intermediate (3.03 g, 91% yield).

Preparation 85

Methyl [4-(4-Hydroxybutyl)phenyl]acetate

A stirred solution of the product of Preparation 84 (2.8 g, 12.8 mmol)in methanol (50 mL) was flushed with nitrogen and then 10% palladium oncarbon (400 mg, 20% wt/wt) was added. The reaction flask was thenalternately placed under vacuum and flushed with hydrogen for cycles andthen stirred under hydrogen for 14 h. The reaction mixture was flushedwith nitrogen and then filtered and the solvent removed under reducedpressure to give the title compound (2.75 g, 97% yield), which was usedwithout further purification.

Preparation 86

Methyl(4-{4-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]butyl}phenyl)acetate(a) Methyl {4-[4-(Toluene-4-sulfonyloxy)butyl]phenyl}acetate

To a stirred solution of the product of Preparation 85 (2.6 g, 12.5mmol) in THF (100 mL) was added DABCO (2.6 g, 25.0 mmol) and thenp-toluenesulfonyl chloride (2.44 g, 13.75 mmol). The reaction mixturewas stirred at room temperature for 23 h and then solvent was removedunder reduced pressure and the residue was dissolved in dichloromethane(200 mL). The organic layer was then washed with water (2×100 mL), 1Nhydrochloric acid (100 mL), aqueous saturated sodium chloride solution(100 mL), dried (MgSO₄), filtered and the solvent removed under reducedpressure to give the title compound, which was used without furtherpurification.

(b) Methyl(4-{4-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]butyl}phenyl)acetate

To the crude product from step (a) was added DMF (50 mL),diisopropylethylamine (3.0 mL, 17.3 mmol) and the product of Preparation8 (2.4 g, 8.1 mmol). The reaction mixture was stirred at roomtemperature for 18 h and then the solvent was removed under reducedpressure to give the title compound (3.5 g, 86.3% yield). MS m/z 501.6(MH⁺), R_(f) 4.89 min (10-70% ACN: H₂O, reverse phase HPLC).

Preparation 87

Biphenyl-2-ylcarbamic Acid1-{4-[4-(2-Hydroxyethyl)phenyl]butyl}piperidin-4-yl Ester

To a stirred solution of the product of Preparation 86 (2.0 g, 4.0 mmol)in THF (100 mL) was added dropwise DIBAL (24 mL, 24 mmol, 1.0 M in THF).After the addition was complete, the reaction mixture was stirred for 3h and then quenched by slow addition of methanol (until gas evolutionceased). The mixture was then stirred for 30 min. and then ethyl acetate(200 mL) and aqueous 1N sodium hydroxide (200 mL) were added. Theorganic layer was separated and washed with aqueous saturated sodiumchloride solution (100 mL), dried (MgSO₄), filtered and the solventremoved under reduced pressure to give the title compound (1.3 g, 69%yield), which was used without further purification. MS m/z 473.4 (MH⁺),R_(f) 4.53 min (10-70% ACN: H₂O, reverse phase HPLC).

Example 23 Biphenyl-2-ylcarbamic Acid1-[2-(4-{2-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)ethyl]piperidin-4-ylEster

To a stirred solution of the product of Preparation 87 (500 mg, 1.06mmol) in dichloromethane (25 mL) was added dimethyl sulfoxide (0.60 mL,10.6 mmol) and diisopropylethylamine (0.921 mL, 5.3 mmol). The reactionmixture was then cooled to −10° C. and pyridine sulfur trioxide (842 mg,5.3 mmol) was added. The reaction mixture was stirred for 1 h and thenquenched by adding water (100 mL). This mixture was stirred for 10 minand then the organic layer was removed and washed with aqueous saturatedsodium chloride solution (100 mL), dried (MgSO₄) and then filtered.

To the filtrate was added methanol (25 mL),5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-oneacetate (419 mg, 1.06 mmol) and sodium triacetoxyborohydride (468 mg,2.12 mmol). This mixture was stirred for 16 h and then condensed and, tothe resulting mixture, was added a 1:1 mixture of acetonitrile andaqueous 4N hydrochloric acid (20 mL). This mixture was heated at 50° C.for 17 h and then the solvent was removed under reduced pressure. To theresidue was added a 1:1 mixture of acetic acid and water (8.0 mL) andthe mixture was chromatographed on reverse-phase silica gel (gradientelution, 10-50% ACN/H₂O) to afford the title compound (67 mg, 7% yieldover 3 steps). MS m/z (MH⁺) 675.5; R_(f) 3.07 (10-70% ACN: H₂O, reversephase HPLC).

Preparation 88

Ethyl 3-[5-(2-Ethoxycarbonylvinyl)thiophen-2-yl]acrylate

To a stirred solution of sodium hydride (2.1 g, 53 mmol, 60% in mineraloil) in THF (200 mL) was slowly added triethylphosphonoacetate (10 mL,50 mmol) Hydrogen gas evolution was observed and the reaction wasstirred until gas evolution ceased (about 30 min). To this reactionmixture was added 2,5-thiophenedicarboxaldehyde (3 g, 21 mmol) and thereaction mixture was stirred for 1 h. The solvent was removed underreduced pressure and the residue was dissolved in dichloromethane (200mL). The organic layer was washed with water (100 mL), aqueous 1Nhydrochloric acid (100 mL), aqueous saturated sodium chloride solution(100 mL), dried (MgSO₄), filtered and the solvent removed under reducedpressure to give the title compound (5.8 g, 98% yield), which was usedwithout further purification.

Preparation 89

Ethyl 3-[5-(2-Ethoxycarbonylethyl)thiophen-2-yl]propionate

A stirred solution of the product of Preparation 88 (5.8 g, 21 mmol) inmethanol (200 mL) was flushed with nitrogen and 10% palladium on carbon(576 mg, 10% wt/wt) was added. The reaction flask was alternately placedunder vacuum and flushed with hydrogen for 3 cycles and then thereaction mixture was stirred under hydrogen for 1 h. The mixture wasthen was flushed with nitrogen, filtered and the solvent removed underreduced pressure to give the title compound (5.8 g, 99% yield), whichwas used without further purification.

Preparation 90

3-[5-(3-Hydroxypropyl)thiophen-2-yl]propan-1-ol

To a stirred solution of DIBAL (88 mL, 88 mmol, 11.0M in cyclohexane) inTHF (300 mL) at −78° C. was added dropwise the product of Preparation 89(5.0 g, 17.6 mmol). After the addition was complete, the reactionmixture was warmed to room temperature over 30 min and then quenched byslow addition of aqueous 1N hydrochloric acid (200 mL). Dichloromethane(400 mL) was added and the layers were separated. The aqueous layer waswashed with dichloromethane (4×100 mL) and the combined organic layerswere washed with aqueous saturated sodium chloride solution (100 mL),dried (MgSO₄), filtered and the solvent removed under reduced pressureto give the title compound (3.0 g, 85% yield), which was used withoutfurther purification.

Preparation 91

Biphenyl-2-ylcarbamic Acid1-{3-[5-(3-Hydroxypropyl)thiophen-2-yl]propyl}piperidin-4-yl Ester (a)Toluene-4-sulfonic Acid 3-[5-(3-Hydroxypropyl)thiophen-2-yl]propyl Ester

To a stirred solution of the product of Preparation 90 (423 mg, 2.1mmol) in THF (20 mL) was added DABCO (420 mg, 4.2 mmol) and thenp-toluenesulfonyl chloride (442 mg, 2.3 mmol). The reaction mixture wasstirred at room temperature for 2 h and then the solvent was removedunder reduced pressure and the residue was dissolved in dichloromethane(200 mL). The organic layer was washed with water (2×100 mL), aqueoussaturated sodium chloride solution (100 mL), dried MgSO₄), filtered andthe solvent removed under reduced pressure to give the title compound,which was used without further purification.

(b) Biphenyl-2-ylcarbamic Acid1-{3-[5-(3-Hydroxypropyl)thiophen-2-yl]propyl}piperidin-4-yl Ester

To the product from step (a) was added acetonitrile (20 mL),diisopropylethylamine (0.5 mL, 2.8 mmol) and the product of Preparation8 (626 mg, 2.11 mmol). The reaction mixture was heated to 50° C. for 20h and then cooled to room temperature and the solvent was removed underreduced pressure. The residue was purified by silica gel chromatography(5% MeOH/DCM with 0.6% NH₃ (aq)) to afford the title compound (450 mg,44% yield). MS m/z (MH⁺) 479.6; R_(f) 4.15 min (10-70% ACN: H₂O, reversephase HPLC).

Preparation 92

Biphenyl-2-ylcarbamic Acid1-[3-(5-{3-[(R)-2-(tert-Butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}thiophen-2-yl)propyl]-piperidin-4-ylEster

To a stirred solution of the product of Preparation 91 (450 mg, 0.94mmol) in dichloromethane (20 mL) was added dimethyl sulfoxide (0.21 mL,3.7 mmol) and diisopropylethylamine (0.65 mL, 3.7 mmol). This mixturewas cooled to −10° C. and pyridine sulfur trioxide (444 mg, 2.8 mmol)was added. The reaction mixture was stirred for 3 h and then wasquenched by adding water (100 mL). This mixture was stirred for 10 minand then the organic layer was removed and was washed with aqueoussaturated sodium chloride solution (100 mL), dried (MgSO₄) and filtered.

To the filtrate was added methanol (20 mL),5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-oneacetate (368 mg, 0.93 mmol) and then sodium triacetoxyborohydride (412mg, 1.86 mmol). This mixture was stirred for 19 h and then the mixturewas condensed to give the title compound, which was used without furtherpurification. MS m/z (MH⁺) 795.8; R_(f) 4.93 min (10-70% ACN: H₂O,reverse phase HPLC).

Example 24 Biphenyl-2-ylcarbamic Acid1-[3-(5-{3-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}thiophen-2-yl)propyl]piperidin-4-ylEster

To the crude product from Preparation 92 was added a 1:1 mixture ofacetonitrile and aqueous 4 N hydrochloric acid (25 mL). This mixture washeated at 50° C. for 17 h and then the solvent was then removed underreduced pressure. To the residue was added a 1:1 mixture of acetic acidand water (8.0 mL) and this mixture was chromatographed on reverse-phasesilica gel (gradient elution, 10-50% ACN/H₂O) to afford the titlecompound (135 mg, 16% yield for 3 steps). MS m/z (MH⁺) 681.5; R_(f) 3.03(10-70% ACN: H₂O, reverse phase HPLC).

Preparation 93

Methyl 4-Amino-5-chloro-2-methoxybenzoate

To a solution of 4-amino-5-chloro-3-methoxybenzoic acid (1.008 g, 5.0mmol) in a mixture of toluene (9 mL) and methanol (1 mL) at 0° C. wasadded (trimethylsilyl)diazomethane (2.0 M in hexane, 3.0 mL, 6.0 mmol)dropwise. The reaction mixture was then warmed to room temperature andstirred for 16 h. Excess (trimethylsilyl)diazomethane was quenched byadding acetic acid until the bright yellow color of the reaction mixturedisappeared. The mixture was then concentrated in vacuo to give thetitle compound as an off-white solid, which was used without furtherpurification.

Preparation 94

Methyl 4-Acryloylamino-5-chloro-2-methoxybenzoate

To crude product of Preparation 93 was added dichloromethane (10 mL, 0.5M) and triethylamine (2.1 mL, 15 mmol). This mixture was cooled to 0° C.and acryloyl chloride (812 μL, 10 mmol) was added dropwise withstirring. After 2 h, the reaction was quenched by adding methanol (about2 mL) at 0° C. and the resulting mixture was stirred at room temperaturefor 15 min and then concentrated in vacuo. Dichloromethane (30 mL) andwater (30 mL) were added to the residue and this mixture was mixedthoroughly. The layers were separated and the aqueous layer wasextracted with dichloromethane (20 mL). The organic layers werecombined, dried (Na₂SO₄), filtered and the solvent was removed in vacuoto give the title compound as a brown foamy solid, which was usedwithout further purification.

Preparation 95

Methyl4-{3-[4-(Biphenyl-2-ylcarbamoyloxy)piperidin-1-yl]propionylamino}-5-chloro-2-methoxybenzoate

To the crude product from Preparation 94 was added the product ofPreparation 8 (1.33 g, 4.5 mmol) and a mixture of THF (22.5 mL) andmethanol (2.5 mL). This mixture was heated at 50° C. with stirring for16 h and then the solvent was removed in vacuo. The residue waschromatographed (silica gel; EtOAc) to give the title compound (0.82 g;R_(f)=0.4, 29% yield over 3 steps) as an off-white foamy solid. MS m/z566.4 (M+H, expected 565.20 for C₃₀H₃₂ClN₃O₆).

Preparation 96

Biphenyl-2-ylcarbamic Acid1-[2-(2-Chloro-4-hydroxymethyl-5-methoxy-phenylcarbamoyl)ethyl]piperidin-4-ylEster

To a solution of the product of Preparation 95 (0.82 mg, 1.45 mmol) in amixture of THF (4.5 mL) and methanol (0.5 mL) at 0° C. was added lithiumborohydride (32 mg, 1.45 mmol). The reaction mixture was allowed to warmto room temperature and was stirred for 41 h. The reaction was thenquenched by adding 1N aqueous hydrochloric acid at 0° C. until no morebubbling was observed and this mixture was stirred for 10 min. Thesolvent was removed in vacuo and the residue was dissolved inacetonitrile (about 2 mL). This solution was purified by prep-RP-HPLC(gradient: 2 to 50% acetonitrile in water with 0.05% TFA). Theappropriate fractions were collected and combined and lyophilized togive the title compound as a trifluoroacetate salt. This salt wastreated with isopropyl acetate (10 mL) and 1N aqueous sodium hydroxide(10 mL) and the organic layer was collected, dried (Na₂SO₄), filteredand the solvent was removed in vacuo to give the title compound (161 mg,21% yield) as a white foamy solid. MS m/z 538.4 (M+H, expected 537.20for C₂₉H₃₂ClN₃O₅).

Preparation 97

Biphenyl-2-ylcarbamic Acid1-[2-(2-Chloro-4-formyl-5-methoxyphenylcarbamoyl)-ethyl]piperidin-4-ylEster

To a solution of the product of Preparation 96 (161 mg, 0.3 mmol) indichloromethane (3 mL) was added dimethyl sulfoxide (213 μL, 3.0 mmol)and diisopropylethylamine (261 μL, 1.5 mmol). This mixture was cooled to−20° C. and sulfur trioxide pyridine complex (238 mg, 1.5 mmol) wasadded slowly. After 30 min, the reaction mixture was quenched by addingwater (about 3 mL). The layers were separated and the organic layer wasdried (Na₂SO₄), filtered and the solvent was removed in vacuo to givethe title compound as a light yellow solid. MS m/z 536.3 (M+H, expected535.19 for C₂₉H₃₀ClN₃O₅).

Preparation 98

Biphenyl-2-ylcarbamic Acid1-[2-(4-([(R)-2-(tert-Butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl)-2-chloro-5-methoxy-phenylcarbamoyl)ethyl]piperidin-4-ylEster

To the product from Preparation 97 in a mixture of dichloromethane (0.5mL) and methanol (0.5 mL) was added5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-oneacetate (124.1 mg, 3.1 mmol) and the resulting mixture was stirred atroom temperature for 1.5 h. Sodium triacetoxyborohydride (190.7 mg, 0.9mmol) was added and the resulting mixture was stirred at roomtemperature for 15 h. The reaction was quenched by adding water (about0.2 mL) and the mixture was concentrated in vacuo to give the titlecompound, which was used without further purification. MS m/z 854.5(M+H, expected 853.36 for C₄₆H₅₆ClN₅O₇Si).

Example 25 Biphenyl-2-ylcarbamic Acid1-[2-(2-Chloro-4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-5-methoxyphenylcarbamoyl)ethyl]piperidin-4-ylEster Ditrifluoroacetate

To a suspension of the product of Preparation 98 in dichloromethane (1.0mL, 0.3 M) was added triethylamine trihydrofluoride (245 μL, 1.5 mmol).This mixture was stirred at room temperature for 45 h and then themixture was concentrated in vacuo. The residue was dissolved in amixture of DMF (0.5 mL), acetonitrile/water (1:1, with 0.1% TFA, 0.6mL), TFA (0.3 mL) and acetonitrile (about 1 mL) and this mixture waspurified by prep-RP-HPLC (gradient: 2 to 50% acetonitrile in water with0.05% TFA). The appropriate fractions were collected and combined andlyophilized to give the title compound (100 mg, 34% yield, 98.7% pure byHPLC) as an off-white solid. MS m/z 740.5 (M+H, expected 739.28 forC₄₀H₄₂ClN₅O₇).

Using the methods described above and the appropriate startingmaterials, the following compounds were prepared. Ex. Compound MS 26Biphenyl-2-ylcarbamic acid 1-{7-[(R)-2-hydroxy-2-(8-hydroxy-2- 613.5oxo-1,2-dihydroquinolin-5-yl)ethylamino]heptyl}piperidin-4-yl ester 27Biphenyl-2-ylcarbamic acid 1-{8-[(R)-2-hydroxy-2-(8-hydroxy-2- 627.5oxo-1,2-dihydroquinolin-5-yl)ethylamino]octyl}piperidin-4-yl ester 28Biphenyl-2-ylcarbamic acid 1-{2-[3-(4-{2-[(R)-2-hydroxy-2-(8- 705.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)ureido]ethyl}piperidin-4-yl ester 29Biphenyl-2-ylcarbamic acid 1-[3-(4-{2-[(R)-2-hydroxy-2-(8- 682.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}piperidin-1-yl)-3-oxopropyl]piperidin-4-yl ester 30Biphenyl-2-ylcarbamic acid 1-{2-[(4-{[(R)-2-hydroxy-2-(8- 682.7hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-cyclohexanecarbonyl)amino]ethyl}piperidin-4-yl ester 31Biphenyl-2-ylcarbamic acid 1-[2-({(1R,3S)-3-[(R)-2-(3- 630.2formylamino-4-hydroxyphenyl)-2-hydroxyethylamino]-cyclopentanecarbonyl}amino)ethyl]piperidin-4-yl ester 32Biphenyl-2-ylcarbamic acid 1-[2-(3-{5-[(R)-2-(3-formylamino-4- 647.5hydroxyphenyl)-2- hydroxyethylamino]pentyl}ureido)ethyl]piperidin-4-ylester 33 Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2-hydroxy-2-(8-662.5 hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)ethyl]piperidin-4-yl ester 34Biphenyl-2-ylcarbamic acid 1-[3-(3-{5-[2-(3-formylamino-4- 661.3hydroxyphenyl)-2- hydroxyethylamino]pentyl}ureido)propyl]piperidin-4-ylester 35 Biphenyl-2-ylcarbamic acid 1-{2-[(4-{2-[(R)-2-hydroxy-2-(8-697.5 hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}piperidine-1-carbonyl)amino]ethyl}piperidin-4-ylester 36 Biphenyl-2-ylcarbamic acid 1-[4-(4-{2-[(R)-2-hydroxy-2-(8-724.5 hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}-phenylamino)benzyl]piperidin-4-yl ester 37 Biphenyl-2-ylcarbamic acid1-[2-(3-{[(R)-2-hydroxy-2-(8-hydroxy- 690.32-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzylcarbamoyl)ethyl]piperidin-4-yl ester 383-[4-(3-Biphenyl-2-yl-ureido)piperidin-1-yl]-N-(4-{[(R)-2-hydroxy- 675.52-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenyl)propionamide 39 Biphenyl-2-ylcarbamic acid1-{2-[(6-{[(R)-2-hydroxy-2-(8- 691.5hydroxy-2-oxo-1,2-dihydroquinolin-5-1)ethylamino]methyl}pyridin-2-ylmethyl)carbamoyl]ethyl}piperidin-4-yl ester 40 Biphenyl-2-ylcarbamicacid 1-[2-(4-{[(R)-2-hydroxy-2-(8-hydroxy- 682.72-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-cyclohexylcarbamoyl)ethyl]piperidin-4-yl ester 41 Biphenyl-2-ylcarbamicacid 1-[2-(4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo- 682.71,2-dihydroquinolin-5-yl)ethylamino]methyl}-cyclohexylcarbamoyl)ethyl]piperidin-4-yl ester 42 Biphenyl-2-ylcarbamicacid 1-[2-({(1R,3S)-3-[(R)-2-hydroxy-2-(8- 654.8hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]-cyclopentanecarbonyl}amino)ethyl]piperidin-4-yl ester 43Biphenyl-2-ylcarbamic acid 1-{2-[(3-{[(R)-2-hydroxy-2-(8- 690.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoyl)methylamino]ethyl}piperidin-4-yl ester 44 Biphenyl-2-ylcarbamicacid 1-{2-[(4-{[(R)-2-hydroxy-2-(8- 696.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-cyclohexanecarbonyl)methylamino]ethyl}piperidin-4-yl ester 45Biphenyl-2-ylcarbamic acid 1-[2-(4-{[2-hydroxy-2-(8-hydroxy-2- NAoxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 46Biphenyl-2-ylcarbamic acid 1-[2-(4-{(S)-1-[(R)-2-hydroxy-2-(8- 690.7hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 47Biphenyl-2-ylcarbamic acid 1-[2-(4-{(R)-1-[(R)-2-hydroxy-2-(8- 690.7hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 48Biphenyl-2-ylcarbamic acid 1-((S)-1-{5-[(R)-2-hydroxy-2-(8- 682.7hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentanoyl}pyrrolidin-2-ylmethyl)piperidin-4-yl ester 49Biphenyl-2-ylcarbamic acid 1-[(S)-1-(4-{[(R)-2-hydroxy-2-(8- 716.8hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoyl)pyrrolidin-2-ylmethyl]piperidin-4-yl ester 50Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-(3-formylamino-4- 652.6hydroxy-phenyl)-2-hydroxy-ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 51Biphenyl-2-ylcarbamic acid 1-[2-(4-{(R)-1-[(R)-2-(3-formylamino- 666.54-hydroxy-phenyl)-2-hydroxy-ethylamino]ethyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 52Biphenyl-2-ylcarbamic acid 1-[2-(4-chloro-3-{[(R)-2-hydroxy-2-(8- 710.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 53N-{2-[4-(3-Biphenyl-2-yl-ureido)-piperidin-1-yl]ethyl}-4-{[(R)-2- 675.5hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzamide 541-Biphenyl-2-yl-3-{1-[3-(4-{2-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo- 681.71,2-dihydroquinolin-5-yl)ethylamino]ethyl}piperidin-1-yl)-3-oxo-propyl]piperidin-4-yl}urea 553-[4-(3-Biphenyl-2-yl-ureido)piperidin-1-yl]-N-(3-{[(R)-2-hydroxy- 689.52-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzyl)propionamide 56 Biphenyl-2-ylcarbamic acid1-(2-fluoro-3-{[(R)-2-hydroxy-2-(8- 637.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzyl)piperidin-4-yl ester 57 Biphenyl-2-ylcarbamic acid1-[2-(3-{[(R)-2-hydroxy-2-(8-hydroxy- 690.42-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-4-methyl-phenylcarbamoyl)ethyl]piperidin-4-yl ester 58 Biphenyl-2-ylcarbamic acid1-[2-(2-chloro-5-{[(R)-2-hydroxy-2-(8- 710.6hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 59Biphenyl-2-ylcarbamic acid 1-[2-(2,6-dichloro-4-{[(R)-2-hydroxy- 745.22-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 60Biphenyl-2-ylcarbamic acid 1-[1-(4-{[(R)-2-hydroxy-2-(8-hydroxy- 730.82-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzoyl)-piperidin-4-ylmethyl]piperidin-4-yl ester 61 Biphenyl-2-ylcarbamic acid1-[2-(4-{[(R)-2-(3-formylamino-4- 652.5hydroxy-phenyl)-2-hydroxyethylamino]methyl}-benzoylamino)ethyl]piperidin-4-yl ester 62 Biphenyl-2-ylcarbamic acid1-{2-[ethyl-(4-{[(R)-2-hydroxy-2-(8- 704.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenyl)carbamoyl]ethyl}piperidin-4-yl ester 63Biphenyl-2-ylcarbamic acid 1-(3-{4-[(R)-2-hydroxy-2-(8-hydroxy- NA2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]piperidin-1-yl}-3-oxo-propyl)piperidin-4-yl ester 64 Biphenyl-2-ylcarbamic acid1-[2-(4-{2-[(R)-2-hydroxy-2-(8- 690.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 65Biphenyl-2-ylcarbamic acid 1-{2-[(5-{[(R)-2-hydroxy-2-(8- 682.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-thiophene-2-carbonyl)amino]ethyl}piperidin-4-yl ester 66Biphenyl-2-ylcarbamic acid 1-{2-[(4-{[(R)-2-hydroxy-2-(8- 735.7hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-3-nitro-benzoyl)methylamino]ethyl}piperidin-4-yl ester 67Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-(3-formylamino-4- 658.8hydroxyphenyl)-2-hydroxyethylamino]methyl}-cyclohexylcarbamoyl)ethyl]piperidin-4-yl ester 68 Biphenyl-2-ylcarbamicacid 1-[2-({4-[(R)-2-hydroxy-2-(8-hydroxy- 682.72-oxo-1,2-dihydroquinolin-5-yl)ethylamino]cyclohexanecarbonyl}-methylamino)ethyl]piperidin-4-yl ester 69 Biphenyl-2-ylcarbamic acid1-(2-fluoro-3-{4-[(R)-2-hydroxy-2-(8- 720.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]piperidin-1-ylmethyl}benzyl)piperidin-4-yl ester 70 Biphenyl-2-ylcarbamic acid1-{2-[(6-{[(R)-2-hydroxy-2-(8- 677.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}pyridine-3-carbonyl)amino]ethyl}piperidin- 4-ylester 71 Biphenyl-2-ylcarbamic acid1-[3-(4-{[(R)-2-hydroxy-2-(8-hydroxy- 654.52-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}piperidin-1-yl)-propyl]piperidin-4-yl ester 72 Biphenyl-2-ylcarbamic acid1-[2-(4-{2-[(R)-2-(3-formylamino-4- 666.5 hydroxyphenyl)-2-hydroxy-ethylamino]ethyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 73Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2-hydroxy-2-(8- 690.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)benzyl]piperidin-4-yl ester 74Biphenyl-2-ylcarbamic acid 1-[2-fluoro-3-(4-{2-[(R)-2-hydroxy-2- 748.5(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}piperidin-1-ylmethyl)benzyl]piperidin-4-yl ester 75Biphenyl-2-ylcarbamic acid 1-[3-(4-{2-[(R)-2-hydroxy-2-(8- 676.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)propyl]piperidin-4-yl ester 76Biphenyl-2-ylcarbamic acid 1-[2-(3-chloro-4-{[(R)-2-hydroxy-2-(8- 710.2hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 77Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2-(8-hydroxy- 769.22-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-2-trifluoromethoxy-phenylcarbamoyl)ethyl]piperidin-4-yl ester 78Biphenyl-2-ylcarbamic acid 1-{3-[3-(4-{2-[(R)-2-hydroxy-2-(8- 752.6hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)phenyl]propyl}piperidin-4-yl ester 79Biphenyl-2-ylcarbamic acid 1-[3-(4-{2-[(S)-2-hydroxy-2-(8- NAhydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)benzyl]piperidin-4-yl ester 80Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2-(8-hydroxy- 802.12-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-2-iodo-phenylcarbamoyl)ethyl]piperidin-4-yl ester 81 Biphenyl-2-ylcarbamic acid1-[2-(2-chloro-4-{[(R)-2-hydroxy-2-(8- 724.2hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-6-methylphenylcarbamoyl)ethyl]piperidin-4-yl ester 82Biphenyl-2-ylcarbamic acid 1-(2-{5-[2-hydroxy-2-(8-hydroxy-2- 656.5oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylcarbamoyl}ethyl)-piperidin-4-yl ester 83 Biphenyl-2-ylcarbamic acid1-[2-(2-bromo-4-{[(R)-2-hydroxy-2-(8- 756.2hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 84Biphenyl-2-ylcarbamic acid 1-{3-[2-(4-{2-[(R)-2-hydroxy-2-(8- 752.8hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)-phenyl]propyl}piperidin-4-yl ester 85Biphenyl-2-ylcarbamic acid 1-[2-fluoro-3-(4-{3-[(R)-2-hydroxy-2- 762.8(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}piperidin-1-ylmethyl)benzyl]piperidin-4-yl ester 86Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2-(8-hydroxy- 706.32-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-2-methoxy-phenylcarbamoyl)ethyl]piperidin-4-yl ester 87 Biphenyl-2-ylcarbamic acid1-[5-(4-{2-[(R)-2-hydroxy-2-(8- 704.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)pentyl]piperidin-4-yl ester 88Biphenyl-2-ylcarbamic acid 1-{2-[1-(4-{2-[(R)-2-hydroxy-2-(8- 730.8hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)-piperidin-4-yl]ethyl}piperidin-4-yl ester 89 Biphenyl-2-ylcarbamic acid1-[2-(4-{2-[(R)-2-hydroxy-2-(8- 704.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]-1-methyl-ethyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 90Biphenyl-2-ylcarbamic acid 1-{2-[4-(4-{2-[(R)-2-hydroxy-2-(8- 744.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)cyclohexyl]ethyl}piperidin-4-yl ester 91Biphenyl-2-ylcarbamic acid 1-[2-(2-fluoro-4-{[(R)-2-hydroxy-2-(8- 694.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 92Biphenyl-2-ylcarbamic acid 1-{2-[3-(4-{2-[(R)-2-hydroxy-2-(8- 738.8hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)phenyl]ethyl}piperidin-4-yl ester 93Biphenyl-2-ylcarbamic acid 1-[2-(2,5-difluoro-4-{[(R)-2-hydroxy-2- 712.3(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 94Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2-hydroxy-2-(8- 690.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}-benzoylamino)ethyl]piperidin-4-yl ester 95 Biphenyl-2-ylcarbamic acid1-[6-(4-{[(R)-2-hydroxy-2-(8-hydroxy- 717.52-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}piperidin-1-ylmethyl)pyridin-2-ylmethyl]piperidin-4-yl ester 96Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2-hydroxy-2-(8- 740.6hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}-naphthalen-1-ylcarbamoyl)ethyl]piperidin-4-yl ester 97Biphenyl-2-ylcarbamic acid 1-{2-[1-(4-{[(R)-2-hydroxy-2-(8- 744.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoyl)piperidin-4-yl]ethyl}piperidin-4-yl ester 98Biphenyl-2-ylcarbamic acid 1-[3-(4-{3-[(R)-2-hydroxy-2-(8- 704.2hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propionylamino}phenyl)propyl]piperidin-4-yl ester 99Biphenyl-2-ylcarbamic acid 1-[3-(4-{[(R)-2-hydroxy-2-(8-hydroxy- 663.72-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)-propyl]piperidin-4-yl ester 100 Biphenyl-2-ylcarbamic acid1-[2-(5-{[(R)-2-hydroxy-2-(8-hydroxy- 673.72-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-1H-benzoimidazol-2-yl)ethyl]piperidin-4-yl ester 101 Biphenyl-2-ylcarbamicacid 1-[2-(4-{3-[(R)-2-hydroxy-2-(8- 696.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propionylamino}cyclohexyl)ethyl]piperidin-4-yl ester 102Biphenyl-2-ylcarbamic acid 1-[2-(4-{5-[(R)-2-hydroxy-2-(8- 724.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentanoylamino}cyclohexyl)ethyl]piperidin-4-yl ester 103Biphenyl-2-ylcarbamic acid 1-[2-(4-{6-[(R)-2-hydroxy-2-(8- 738.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]-hexanoylamino}cyclohexyl)ethyl]piperidin-4-yl ester 104Biphenyl-2-ylcarbamic acid 1-[2-(1-{3-[(R)-2-hydroxy-2-(8- 682.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propionyl}piperidin-4-yl)ethyl]piperidin-4-yl ester 105Biphenyl-2-ylcarbamic acid 1-{2-[3-(4-{[(R)-2-hydroxy-2-(8- 691.7hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenyl)ureido]ethyl}piperidin-4-yl ester 106Biphenyl-2-ylcarbamic acid 1-{2-[(2-{4-[(R)-2-hydroxy-2-(8- 682.7hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]cyclohexyl}-ethyl)methylamino]ethyl}piperidin-4-yl ester 107 Biphenyl-2-ylcarbamicacid 1-[2-(2,3,5,6-tetrafluoro-4-{[(R)-2- 748.2hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]piperidin-4-yl ester 108Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2-(8-hydroxy- 928.02-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-2,6-diiodo-phenylcarbamoyl)ethyl]piperidin-4-yl ester 109 Biphenyl-2-ylcarbamicacid 1-[2-(1-{4-[(R)-2-hydroxy-2-(8- 696.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]-butyryl}piperidin-4-yl)ethyl]piperidin-4-yl ester 110Biphenyl-2-ylcarbamic acid 1-[2-(1-{5-[(R)-2-hydroxy-2-(8- 710.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentanoyl}piperidin-4-yl)ethyl]piperidin-4-yl ester 111Biphenyl-2-ylcarbamic acid 1-[2-(1-{6-[(R)-2-hydroxy-2-(8- 724.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]-hexanoyl}piperidin-4-yl)ethyl]piperidin-4-yl ester 112Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-hydroxy-2-(8-hydroxy- 690.52-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzylcarbamoyl)ethyl]piperidin-4-yl ester 113 Biphenyl-2-ylcarbamicacid 1-[2-(4-{[(R)-2-(3-formylamino-4- 666.5hydroxy-phenyl)-2-hydroxyethylamino]methyl}-benzylcarbamoyl)ethyl]piperidin-4-yl ester 114 Biphenyl-2-ylcarbamicacid 1-{2-[3-(4-{[(R)-2-hydroxy-2-(8- 705.6hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzyl)ureido]ethyl}piperidin-4-yl ester 115 Biphenyl-2-ylcarbamic acid1-{2-[3-(4-{[(R)-2-(3-formylamino-4- 681.7hydroxyphenyl)-2-hydroxyethylamino]methyl}benzyl)-ureido]ethyl}piperidin-4-yl ester 116 Biphenyl-2-ylcarbamic acid1-[2-(4-{[(R)-2-hydroxy-2-(8-hydroxy- 690.42-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-2-methyl-phenylcarbamoyl)ethyl]piperidin-4-yl ester 117 Biphenyl-2-ylcarbamicacid 1-(3-{4-[2-(4-{[(R)-2-hydroxy-2-(8- 774.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}piperidin-1-yl)ethyl]phenoxy}propyl)-piperidin-4-yl ester 118 Biphenyl-2-ylcarbamic acid1-[2-(3-{2-[(R)-2-hydroxy-2-(8- 690.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}-benzylcarbamoyl)ethyl]piperidin-4-yl ester 119 Biphenyl-2-ylcarbamicacid 1-[2-(3-{[(R)-2-hydroxy-2-(8-hydroxy- 649.52-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)ethyl]piperidin-4-yl ester 120Biphenyl-2-ylcarbamic acid 1-(2-{[2-(4-{[(R)-2-hydroxy-2-(8- 720.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl]methylamino}ethyl)- piperidin-4-ylester 121 Biphenyl-2-ylcarbamic acid 1-(2-{[2-(3-{[(R)-2-hydroxy-2-(8-720.4 hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl]methylamino}ethyl)- piperidin-4-ylester 122 Biphenyl-2-ylcarbamic acid 1-{2-[(5-{[(R)-2-hydroxy-2-(8-680.3 hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}furan-2-carbonyl) methylamino]ethyl}piperidin-4-yl ester 123Biphenyl-2-ylcarbamic acid 1-{2-[(5-{[(R)-2-hydroxy-2-(8- 696.2hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-thiophene-2-carbonyl)methylamino]ethyl}piperidin-4-yl ester 124Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2-hydroxy-2-(8- 679.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethoxy}phenoxy)ethyl]piperidin-4-yl ester 125Biphenyl-2-ylcarbamic acid 1-{2-[4-(4-{[(R)-2-hydroxy-2-(8- 758.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoylamino)cyclohexyl]ethyl}piperidin-4-yl ester 126Biphenyl-2-ylcarbamic acid 1-(2-{4-[2-(2-{[(R)-2-hydroxy-2-(8- 788.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetylamino]cyclohexyl}ethyl)-piperidin-4-yl ester 127 Biphenyl-2-ylcarbamic acid1-(2-{4-[2-(3-{[(R)-2-hydroxy-2-(8- 788.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetylamino]cyclohexyl}ethyl)-piperidin-4-yl ester 128 Biphenyl-2-ylcarbamic acid1-(2-{4-[2-(4-{[(R)-2-hydroxy-2-(8- 788.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetylamino]cyclohexyl}ethyl)-piperidin-4-yl ester 129 Biphenyl-2-ylcarbamic acid1-(2-{4-[(5-{[(R)-2-hydroxy-2-(8- 748.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}furan-2-carbonyl)amino]cyclohexyl}ethyl)piperidin-4-yl ester 130Biphenyl-2-ylcarbamic acid 1-(2-{4-[(5-{[(R)-2-hydroxy-2-(8- 764.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-thiophene-2-carbonyl)amino]cyclohexyl}ethyl)piperidin-4-yl ester 131Biphenyl-2-ylcarbamic acid 1-(2-{1-[2-(2-{[(R)-2-hydroxy-2-(8- 774.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl]piperidin-4-yl}ethyl)-piperidin-4-yl ester 132 Biphenyl-2-ylcarbamic acid1-(2-{1-[2-(3-{[(R)-2-hydroxy-2-(8- 774.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl]piperidin-4-yl}ethyl)-piperidin-4-yl ester 133 Biphenyl-2-ylcarbamic acid1-(2-{1-[2-(4-{[(R)-2-hydroxy-2-(8- 774.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetyl]piperidin-4-yl}ethyl)-piperidin-4-yl ester 134 Biphenyl-2-ylcarbamic acid1-{2-[1-(5-{[(R)-2-hydroxy-2-(8- 734.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}furan-2-carbonyl) piperidin-4-yl]ethyl}piperidin-4-yl ester 135Biphenyl-2-ylcarbamic acid 1-{2-[1-(5-{[(R)-2-hydroxy-2-(8- 750.2hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-thiophene-2-carbonyl)piperidin-4-yl]ethyl}piperidin-4-yl ester 136Biphenyl-2-ylcarbamic acid 1-{2-[4-(3-{[(R)-2-hydroxy-2-(8- 752.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoylamino)phenyl]ethyl}piperidin-4-yl ester 137 Biphenyl-2-ylcarbamicacid 1-{2-[4-(4-{[(R)-2-hydroxy-2-(8- 752.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoylamino)phenyl]ethyl}piperidin-4-yl ester 138 Biphenyl-2-ylcarbamicacid 1-(2-{4-[2-(2-{[(R)-2-hydroxy-2-(8- 782.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetylamino]phenyl}ethyl)- piperidin-4-ylester 139 Biphenyl-2-ylcarbamic acid 1-(2-{4-[2-(3-{[(R)-2-hydroxy-2-(8-782.4 hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetylamino]phenyl}ethyl)- piperidin-4-ylester 140 Biphenyl-2-ylcarbamic acid 1-(2-{4-[2-(4-{[(R)-2-hydroxy-2-(8-782.4 hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)acetylamino]phenyl}ethyl)- piperidin-4-ylester 141 Biphenyl-2-ylcarbamic acid 1-(2-{4-[(5-{[(R)-2-hydroxy-2-(8-742.4 hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}furan-2-carbonyl)amino]phenyl}ethyl)piperidin-4-yl ester 142Biphenyl-2-ylcarbamic acid 1-(2-{4-[(5-{[(R)-2-hydroxy-2-(8- 758.2hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-thiophene-2-carbonyl)amino]phenyl}ethyl)piperidin-4-yl ester 143Biphenyl-2-ylcarbamic acid 1-{2-[4-(3-{[(R)-2-hydroxy-2-(8- 758.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoylamino)cyclohexyl]ethyl}piperidin-4-yl ester 144Biphenyl-2-ylcarbamic acid 1-[3-(3-{[(R)-2-hydroxy-2-(8-hydroxy- 663.42-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)-propyl]piperidin-4-yl ester 145 Biphenyl-2-ylcarbamic acid1-[2-hydroxy-3-(4-{2-[(R)-2-hydroxy- 692.32-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)propyl]piperidin-4-yl ester 146Biphenyl-2-ylcarbamic acid 1-[4-(4-{2-[(R)-2-hydroxy-2-(8- 690.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)butyl]piperidin-4-yl ester 147Biphenyl-2-ylcarbamic acid 1-{2-[4-({2-[(R)-2-hydroxy-2-(8- 733.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]acetylamino}-methyl)phenylcarbamoyl]ethyl}piperidin-4-yl ester 148Biphenyl-2-ylcarbamic acid 1-{2-[4-(2-{2-[(R)-2-hydroxy-2-(8- 747.4hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]acetylamino}-ethyl)phenylcarbamoyl]ethyl}piperidin-4-yl ester 149Biphenyl-2-ylcarbamic acid 1-{2-[(4-{[(R)-2-hydroxy-2-(8- 696.6hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-cyclohexylmethyl)carbamoyl]ethyl}piperidin-4-yl ester 150Biphenyl-2-ylcarbamic acid 1-(2-{6-[(R)-2-hydroxy-2-(8-hydroxy- 656.62-oxo-1,2-dihydroquinolin-5-yl)ethylamino]hexanoylamino}-ethyl)piperidin-4-yl ester 151 Biphenyl-2-ylcarbamic acid1-[2-(3-{2-[(R)-2-hydroxy-2-(8- 679.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethoxy}phenoxy)ethyl]piperidin-4-yl ester 152Biphenyl-2-ylcarbamic acid 1-[2-(2-{2-[(S)-2-hydroxy-2-(8- 679.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethoxy}phenoxy)ethyl]piperidin-4-yl ester 153Biphenyl-2-ylcarbamic acid 1-[2-(2-{[(R)-2-hydroxy-2-(8-hydroxy- 711.32-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenoxy)benzyl]piperidin-4-yl ester 154Biphenyl-2-ylcarbamic acid 1-(2-{6-[(R)-2-hydroxy-2-(8-hydroxy- 670.42-oxo-1,2-dihydroquinolin-5-yl)ethylamino]hexylcarbamoyl}ethyl)piperidin-4-yl ester 155 Biphenyl-2-ylcarbamic acid1-[2-({(1R,3S)-3-[(R)-2-hydroxy-2-(8- 654.8hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]cyclopentanecarbonyl}amino)ethyl]piperidin-4-yl ester 156Biphenyl-2-ylcarbamic acid 1-[3-(4-{3-[(R)-2-hydroxy-2-(8- 675.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}phenyl)propyl]piperidin-4-yl ester 157Biphenyl-2-ylcarbamic acid 1-[3-(4-{2-[(R)-2-hydroxy-2-(8- 661.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)propyl]piperidin-4-yl ester 158Biphenyl-2-ylcarbamic acid 1-[4-(4-{2-[(R)-2-hydroxy-2-(8- 675.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)butyl]piperidin-4-yl ester 159Biphenyl-2-ylcarbamic acid 1-[3-(5-{3-[(R)-2-hydroxy-2-(8- 665.6hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}furan-2-yl)propyl]piperidin-4-yl ester 160 Biphenyl-2-ylcarbamic acid1-{2-[3-(4-{2-[(R)-2-hydroxy-2-(8- 719.2hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)-1-methylureido]ethyl}piperidin-4-yl ester 161 Biphenyl-2-ylcarbamic acid1-{2-[1-(4-{2-[(R)-2-hydroxy-2-(8- 773.3hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylcarbamoyl)piperidin-4- yl]ethyl}piperidin-4-ylester 162 Biphenyl-2-ylcarbamic acid 1-[3-(3-{3-[(R)-2-hydroxy-2-(8-675.5 hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}phenyl)propyl]piperidin-4-yl ester 163Biphenyl-2-ylcarbamic acid 1-[3-(5-{3-[(R)-2-hydroxy-2-(8- 669.6hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]propyl}tetrahydrofuran-2-yl)propyl]piperidin-4-yl ester164 Biphenyl-2-ylcarbamic acid 1-[2-(4-{2-[(R)-2-hydroxy-2-(8- 706.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethylcarbamoyl}phenoxy)ethyl]piperidin-4-yl ester 165(5-Bromobiphenyl-2-yl)carbamic acid 1-{9-[2-hydroxy-2-(8- NAhydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}- piperidin-4-ylester 166 (2′-Fluorobiphenyl-2-yl)carbamic acid1-{9-[(R)-2-hydroxy-2-(8- 659.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}- piperidin-4-ylester 167 (3′-Chloro-3,5-difluorobiphenyl-2-yl)carbamic acid1-{9-[(R)-2- 711.8 hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)-ethylamino]nonyl}piperidin-4-yl ester 168(3′,5′-Dichloro-3,5-difluorobiphenyl-2-yl)carbamic acid 1-{9-[(R)-2-745.5 hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)-ethylamino]nonyl}piperidin-4-yl ester 169(3,5-Difluorobiphenyl-2-yl)carbamic acid 1-{9-[(R)-2-hydroxy-2-(8- 677.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}- piperidin-4-ylesterPreparation 99

Biphenyl-2-ylcarbamic Acid1-[2-(4-[1,3]dioxolan-2-ylphenylcarbamoyl)-ethyl]-4-methylpiperidin-4-ylEster

A mixture of biphenyl-2-ylcarbamic acid 4-methylpiperidin-4-yl ester(2.73 g, 8.79 mmol) and N-(4-[1,3]dioxolan-2-yl-phenyl)acrylamide (2.05g, 8.80 mmol) were heated in 100 mL of 1:1 methanol/dichloromethane at50° C. under nitrogen for 1 h. The solution was then diluted with ethylacetate and the organic layer was washed with water, brine, dried(MgSO₄) and concentrated under reduced pressure to give the titlecompound. MS m/z calcd for C₃₁H₃₅N₃O₅ (M+H)⁺ 530.6. found 530.4.

Preparation 100

Biphenyl-2-ylcarbamic Acid1-[2-(4-Formylphenylcarbamoyl)ethyl]-4-methylpiperidin-4-yl Ester

The product of Preparation 99 was redissolved in 40 mL of methanol and25 mL of aqueous 1 N hydrochloric acid was added. The resulting mixturewas stirred at room temperature overnight and the organic solvent wasremoved under reduced pressure. The residue was dissolved in ethylacetate and the organic layer was washed with water, brine, dried(MgSO₄) and the solvent removed under reduced pressure. The product wastriturated with dichloromethane to give the title compound as a whitepowder (2.47 g). LCMS (2-90) R_(t)=4.27 min; MS m/z calcd for C₂₉H₃₁N₃O₄(M+H)⁺ 486.6. found 486.5.

Preparation 101

Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-(tert-butyldimethylsilanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]-4-methyl-piperidin-4-ylEster

A mixture of the product of Preparation 100 (1.70 g, 3.51 mmol) and5-[(R)-2-amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-oneacetate (1.65 g, 4.19 mmol) in 40 mL of 1:1 methanol and dichloromethanewas stirred at room temperature overnight. Sodium triacetoxyborohydride(2.23 g, 10.5 mmol) was then added in one portion and the reactionmixture was stirred at room temperature for 6 hr. The reaction was thenquenched with water and diluted with ethyl acetate. The layers wereseparated and the organic layer was washed with saturated sodiumbicarbonate, brine, dried (MgSO₄) and the solvent removed under reducedpressure to give the title compound (2.9 g). MS m/z calcd forC₄₆H₅₇N₅O₆Si (M+H)+ 805.0. found 804.6.

Example 170 Biphenyl-2-ylcarbamic Acid1-[2-(4-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-phenylcarbamoyl)ethyl]-4-methylpiperidin-4-ylEster

The product of Preparation 101 (2.9 g, 3.6 mmol) was dissolved in 75 mLof dichloromethane and triethylamine trihydrofluoride (0.85 mL, 5.2mmol) was added. The resulting mixture was stirred at room temperatureovernight and then the solvent was removed under reduced pressure togive the crude product as an oil. The product was then dissolved inacetic acid/water (1:1) and purified by prep HPLC to give the titlecompound as an off-white solid. LCMS (2-90) R_(t)=3.67 min.; MS m/zcalcd C₄₀H₄₃N₅O₆ (M+H)⁺ 690.8. found 690.3.

Using the methods described herein and the appropriate startingmaterials, the following compounds can be prepared. Ex. Compound MS 171Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-(3-formylamino-4- NAhydroxyphenyl)-2-hydroxyethylamino]methyl}phenylcarbamoyl)-ethyl]-4-methylpiperidin-4-yl ester 172 Biphenyl-2-ylcarbamic acid1-{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo- NA1,2-dihydroquinolin-5-yl)ethylamino]nonyl}-4- methylpiperidin-4-yl ester173 Biphenyl-2-ylcarbamic acid 1-{9-[(R)-2-(3-formylamino-4-hydroxy- NAphenyl)-2-hydroxyethylamino]nonyl}-4-methylpiperidin-4-yl ester 174Biphenyl-2-ylcarbamic acid 1-(2-{5-[(R)-2-hydroxy-2-(8-hydroxy- NA2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]pentylcarbamoyl}-ethyl)-4-methylpiperidin-4-yl ester 175 Biphenyl-2-ylcarbamic acid1-(2-{5-[(R)-2-(3-formylamino-4- NAhydroxyphenyl)-2-hydroxyethylamino]pentylcarbamoyl}ethyl)-4-methylpiperidin-4-yl ester 176 Biphenyl-2-ylcarbamic acid1-(2-{6-[(R)-2-hydroxy-2-(8-hydroxy- NA2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]hexanoylamino}ethyl)-4-methylpiperidin-4-yl ester 177 Biphenyl-2-ylcarbamic acid1-(2-{6-[(R)-2-(3-formylamino-4- NAhydroxyphenyl)-2-hydroxyethylamino]hexanoylamino}ethyl)-4-methylpiperidin-4-yl ester 178 Biphenyl-2-ylcarbamic acid1-[2-(4-{[(R)-2-hydroxy-2-(8-hydroxy- NA2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-benzoylamino)ethyl]-4-methylpiperidin-4-yl ester 179Biphenyl-2-ylcarbamic acid 1-[2-(4-{[(R)-2-(3-formylamino-4- NAhydroxyphenyl)-2-hydroxyethylamino]methyl}benzoylamino)-ethyl]-4-methylpiperidin-4-yl ester 180 Biphenyl-2-ylcarbamic acid1-{3-[4-(4-{2-[(R)-2-hydroxy-2-(8- 776.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenylamino)phenyl]propyl}-4- methylpiperidin-4-ylester 181 Biphenyl-2-ylcarbamic acid1-[2-(2-chloro-4-{[(R)-2-hydroxy-2-(8- 724.5hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}phenylcarbamoyl)ethyl]-4-methylpiperidin-4-yl ester182 Biphenyl-2-ylcarbamic acid 1-[2-(2-chloro-4-{[(R)-2-hydroxy-2-(8-754.5 hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-5-methoxyphenylcarbamoyl)ethyl]-4-methylpiperidin-4-yl esterPreparation 102

Biphenyl-2-ylcarbamic acid (R)-(1-azabicyclo[3.2.1]oct-4-yl)Ester

2-Biphenyl isocyanate (1.00 g, 5.12 mmol) and (R)-(−)-3-quinuclidinolhydrochloride (921 mg, 5.63 mmol) were heated together inN,N-dimethylformamide (2.06 mL) at 110° C. for 12 h. The reactionmixture was cooled and diluted with ethyl acetate (15 mL) and thenwashed with saturated aqueous sodium bicarbonate (2×10 mL). The organiclayer was extracted with 1 M hydrochloric acid (3×20 mL) and thecombined aqueous extracts were made basic to pH 8-9 with potassiumcarbonate. The aqueous layer was then extracted with ethyl acetate (3×20mL) and the combined organic layers were dried (magnesium sulfate) andsolvent was removed under reduced pressure to give the title compound asa yellow oil (1.64 g, 99% yield).

Preparation 103

(R)-4-(Biphenyl-2-ylcarbamoyloxy)-1-(9-bromononyl)-1-azoniabicyclo[3.2.1]octaneBromide

To a stirred solution of the product of Preparation 102 (1.21 g, 3.76mmol) and triethylamine (1.05 mL, 7.52 mmol) in acetonitrile (18.8 mL)was added 1,9-dibromononane (994 μL, 4.89 mmol) and the reaction mixturewas heated at 50° C. for 4 h. The reaction mixture was then cooled andthe solvent was removed under reduced pressure. The residue wasdissolved in dichloromethane (20 mL) and the organic layer was washedwith saturated aqueous sodium bicarbonate (10 mL), dried (magnesiumsulfate) and solvent removed under reduced pressure. The crude productwas purified by flash chromatography (10% methanol/dichloromethane, 0.5%ammonium hydroxide) to give the title compound (1.04 g, 1.97 mmol, 52%yield).

Preparation 104

(R)-1-(9-N,N-Di(tert-butoxycarbonyl)aminononyl)-4-(biphenyl-2-ylcarbamoyloxy)-1-azoniabicyclo[3.2.1]octaneBromide

To a stirred solution of sodium hydride (60% dispersion in mineral oil)(126 mg, 3.15 mmol) in N,N-dimethylformamide (10 mL) under an atmosphereof nitrogen at 0° C., was added di-tert-butyl iminodicarboxylate (513mg, 2.36 mmol) in N,N-dimethylformamide (5 mL). The reaction mixture wasstirred at room temperature for 15 min and then it was cooled to 0° C.and the product of Preparation 103 (1.04 g, 1.97 mmol) inN,N-dimethylformamide (5 mL) was added. The reaction mixture was allowedto warm to room temperature over a 12 h period and then the solvent wasremoved under reduced pressure to give the title compound, which wasused without further purification.

Preparation 105

(R)-1-(9-Aminononyl)-4-(biphenyl-2-ylcarbamoyloxy)-1-azoniabicyclo[3.2.1]octaneBromide

The product of Preparation 104 (1.31 g, 1.97 mmol) was dissolved indichloromethane (15 mL) and trifluoroacetic acid (5 mL) was addedslowly. The reaction mixture was stirred at room temperature for 1 h andthen the solvent was removed under reduced pressure. The residue wasdissolved in dichloromethane (20 mL) and washed with aqueous 1 M sodiumhydroxide (20 mL). The organic layer was extracted with 1 M hydrochloricacid (3×20 mL) and the combined aqueous extracts were made basic withpotassium carbonate and extracted with dichloromethane (3×20 mL). Thecombined organic layers were dried (magnesium sulfate) and solvent wasremoved under reduced pressure to give the title compound (210 mg, 23%yield over 2 steps).

Preparation 106

(R)-1-{9-[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydroquinolin-5-yl)-2-hydroxyethylamino]-nonyl}-4-(biphenyl-2-ylcarbamoyloxy)-1-azoniabicyclo[3.2.1]octaneBromide

The product of Preparation 105 (210 mg, 0.45 mmol) and sodiumtriacetoxyborohydride (286 mg, 1.35 mmol) were stirred in dichloroethane(4.5 mL) at room temperature for 2 h and then the product of Preparation6 (163 mg, 0.50 mmol) was added. The reaction mixture was stirred for 12h and then diluted with dichloromethane (10 mL) and washed withsaturated aqueous sodium bicarbonate (10 mL), dried (magnesium sulfate)and solvent removed under reduced pressure. The crude reaction productwas purified by flash chromatography (10-50% methanol/dichloromethane,0.5% ammonium hydroxide) to give the title compound (131 mg, 38% yield).

Example 1834-(Biphenyl-2-ylcarbamoyloxy)-1-{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)-ethylamino]nonyl}-1-azoniabicyclo[2.2.2]octaneBromide Ditrifluoroacetate

To a stirred solution of the product of Preparation 105 (131 mg, 0.17mmol) in methanol (1.8 mL) was added palladium (10 wt. % dry basis onactivated carbon; 39 mg) and the reaction mixture was placed under anatmosphere of hydrogen. After stirring for 12 h, the reaction mixturewas filtered through a pad of Celite, washing with methanol (2 mL) andsolvent removed under reduced pressure. The resulting residue waspurified by preparative HPLC to give the title compound as itsditrifluoroacetate salt (8 mg). MS m/z 667.5.

Using the methods described herein and the appropriate startingmaterials, the following compounds can be prepared. Ex. Compound MS 184Biphenyl-2-ylcarbamic acid 8-{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-667.3 dihydroquinolin-5-yl)ethylamino]nonyl}-8-aza-bicyclo[3.2.1]oct-3-yl ester 1857-(Biphenyl-2-ylcarbamoyloxy)-9-{9-[(R)-2-hydroxy-2-(8-hydroxy- 695.52-oxo-1,2-dihydroquinolin-5-yl)-ethylamino]nonyl}-9-methyl-3-oxa-9-azoniatricyclo[3.3.1.0*2,4*]nonane bromide 186Biphenyl-2-ylcarbamic acid 9-{9-[(R)-2-hydroxy-2-(8-hydroxy-2- 681.5oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}-3-oxa-9-aza-tricyclo[3.3.1.0*2,4*]non-7-yl esterPreparation aCell Culture and Membrane Preparation from Cells Expressing Human β₁, β₂or β₃ Adrenergic Receptors

Chinese hamster ovarian (CHO) cell lines stably expressing cloned humanβ₁, β₂ or β₃ adrenergic receptors, respectively, were grown to nearconfluency in Hams F-12 media with 10% FBS in the presence of 500 μg/mLGeneticin. The cell monolayer was lifted with 2 mM EDTA in PBS. Cellswere pelleted by centrifugation at 1,000 rpm, and cell pellets wereeither stored frozen at −80° C. or membranes were prepared immediatelyfor use. For preparation of β₁ and β₂ receptor expressing membranes,cell pellets were re-suspended in lysis buffer (10 mM HEPES/HCl, 10 mMEDTA, pH 7.4 at 4° C.) and homogenized using a tight-fitting Dounceglass homogenizer (30 strokes) on ice. For the more protease-sensitiveβ₃ receptor expressing membranes, cell pellets were homogenated in lysisbuffer (10 mM Tris/HCl, pH 7.4) supplemented with one tablet of“Complete Protease Inhibitor Cocktail Tablets with 2 mM EDTA” per 50 mLbuffer (Roche Catalog No. 1697498, Roche Molecular Biochemicals,Indianapolis, Ind.). The homogenate was centrifuged at 20,000×g, and theresulting pellet was washed once with lysis buffer by re-suspension andcentrifugation as above. The final pellet was then re-suspended inice-cold binding assay buffer (75 mM Tris/HCl pH 7.4, 12.5 mM MgCl₂, 1mM EDTA). The protein concentration of the membrane suspension wasdetermined by the methods described in Lowry et al., 1951, Journal ofBiological Chemistry, 193, 265; and Bradford, Analytical Biochemistry,1976, 72, 248-54. All membranes were stored frozen in aliquots at −80°C. or used immediately.

Preparation B

Cell Culture and Membrane Preparation from Cells Expressing Human M₁,M₂, M₃ and M₄ Muscarinic Receptors

CHO cell lines stably expressing cloned human hM₁, hM₂, hM₃ and hM₄muscarinic receptor subtypes, respectively, were grown to nearconfluency in HAM's F-12 media supplemented with 10% FBS and 250 μg/mLGeneticin. The cells were grown in a 5% CO₂, 37° C. incubator and liftedwith 2 mM EDTA in dPBS. Cells were collected by 5 minute centrifugationat 650×g, and cell pellets were either stored frozen at −80° C. ormembranes were prepared immediately for use. For membrane preparation,cell pellets were resuspended in lysis buffer and homogenized with aPolytron PT-2100 tissue disrupter (Kinematica AG; 20 seconds×2 bursts).Crude membranes were centrifuged at 40,000×g for 15 minutes at 4° C. Themembrane pellet was then resuspended with re-suspension buffer andhomogenized again with the Polytron tissue disrupter. The proteinconcentration of the membrane suspension was determined by the methoddescribed in Lowry et al., 1951, Journal of Biochemistry, 193, 265. Allmembranes were stored frozen in aliquots at −80° C. or used immediately.Aliquots of prepared hM₅ receptor membranes were purchased directly fromPerkin Elmer and stored at −80° C. until use.

Assay Test Procedure A

Radioligand Binding Assay for Human β₁, β₂ and β₃ Adrenergic Receptors

Binding assays were performed in 96-well microtiter plates in a totalassay volume of 100 μL with 10-15 μg of membrane protein containing thehuman β₁, β₂ or β₃ adrenergic receptors in assay buffer (75 mM Tris/HClpH 7.4 at 25° C., 12.5 mM MgCl₂, 1 mM EDTA, 0.2% BSA). Saturationbinding studies for determination of K_(d) values of the radioligandwere done using [³H]-dihydroalprenolol (NET-720, 100 Ci/mmol,PerkinElmer Life Sciences Inc., Boston, Mass.) for the β₁ and β₂receptors and [¹²⁵I]-(−)-iodocyanopindolol (NEX-189, 220 Ci/mmol,PerkinElmer Life Sciences Inc., Boston, Mass.) at 10 or 11 differentconcentrations ranging from 0.01 nM to 20 nM. Displacement assays fordetermination of K_(i) values of test compounds were done with[³H]-dihydroalprenolol at 1 nM and [¹²⁵I]-(−)-iodocyanopindolol at 0.5nM for 10 or 11 different concentrations of test compound ranging from10 pM to 10 μM. Non-specific binding was determined in the presence of10 μM propranolol. Assays were incubated for 1 hour at 37° C., and thenbinding reactions were terminated by rapid filtration over GF/B for theβ₁ and β₂ receptors or GF/C glass fiber filter plates for the β₃receptors (Packard BioScience Co., Meriden, Conn.) presoaked in 0.3%polyethyleneimine. Filter plates were washed three times with filtrationbuffer (75 mM Tris/HCl pH 7.4 at 4° C., 12.5 mM MgCl₂, 1 mM EDTA) toremove unbound radioactivity. The plates were then dried and 50 μL ofMicroscint-20 liquid scintillation fluid (Packard BioScience Co.,Meriden, Conn.) was added and plates were counted in a Packard Topcountliquid scintillation counter (Packard BioScience Co., Meriden, Conn.).Binding data were analyzed by nonlinear regression analysis with theGraphPad Prism Software package (GraphPad Software, Inc., San Diego,Calif.) using the 3-parameter model for one-site competition. The curveminimum was fixed to the value for nonspecific binding, as determined inthe presence of 10 μM propranolol. K_(i) values for test compounds werecalculated from observed IC₅₀ values and the K_(d) value of theradioligand using the Cheng-Prusoff equation (Cheng Y, and Prusoff W H.,Biochemical Pharmacology, 1973, 22, 23, 3099-108).

In this assay, a lower K_(i) value indicates that a test compound has ahigher binding affinity for the receptor tested. Exemplified compound ofthis invention that were tested in this assay typically were found tohave a K_(i) value of less than about 300 nM for the β₂ adrenergicreceptor. For example, the compounds of Examples 3 and 6 were found tohave K_(i) values of less than 10 nM.

If desired, the receptor subtype selectivity for a test compound can becalculated as the ratio of K_(i)(β₁)/K_(i)(β₂) or K_(i)(β₃)/K_(i)(β₂).Typically, compounds of this invention demonstrated greater binding atthe β₂ adrenergic receptor compared to the β₁ or β₃ adrenergic receptor,i.e. K_(i)(β₁) or K_(i)(β₃) is typically greater than K_(i)(β₂).Generally, compounds having selectivity for the β₂ adrenergic receptorover the β₁ or β₃ adrenergic receptors are preferred; especiallycompounds having a selectivity greater than about 5; and in particular,greater than about 8. By way of example, the compounds of Examples 3 and6 had a ratio of K_(i)(β₁)/K_(i)(β₂) greater than 8.

Assay Test Procedure B

Radioligand Binding Assay for Muscarinic Receptors

Radioligand binding assays for cloned human muscarinic receptors wereperformed in 96-well microtiter plates in a total assay volume of 100μL. CHO cell membranes stably expressing either the hM₁, hM₂, hM₃, hM₄or hM₅ muscarinic subtype were diluted in assay buffer to the followingspecific target protein concentrations (μg/well): 10 μg for hM₁, 10-15μg for hM₂, 10-20 μg for hM₃, 10-20 μg for hM₄, and 10-12 μg for hM₅ toget similar signals (cpm). The membranes were briefly homogenized usinga Polytron tissue disruptor (10 seconds) prior to assay plate addition.Saturation binding studies for determining K_(D) values of theradioligand were performed using L-[N-methyl-³H]scopolamine methylchloride ([³H]-NMS) (TRK666, 84.0 Ci/mmol, Amersham Pharmacia Biotech,Buckinghamshire, England) at concentrations ranging from 0.001 nM to 20nM. Displacement assays for determination of K_(i) values of testcompounds were performed with [³H]-NMS at 1 nM and eleven different testcompound concentrations. The test compounds were initially dissolved toa concentration of 400 μM in dilution buffer and then serially diluted5× with dilution buffer to final concentrations ranging from 10 pM to100 μM. The addition order and volumes to the assay plates were asfollows: 25 μL radioligand, 25 μL diluted test compound, and 50 μLmembranes. Assay plates were incubated for 60 minutes at 37° C. Bindingreactions were terminated by rapid filtration over GF/B glass fiberfilter plates (PerkinElmer Inc., Wellesley, Mass.) pre-treated in 1%BSA. Filter plates were rinsed three times with wash buffer (10 mMHEPES) to remove unbound radioactivity. The plates were then air driedand 50 μL Microscint-20 liquid scintillation fluid (PerkinElmer Inc.,Wellesley, Mass.) was added to each well. The plates were then countedin a PerkinElmer Topcount liquid scintillation counter (PerkinElmerInc., Wellesley, Mass.). Binding data were analyzed by nonlinearregression analysis with the GraphPad Prism Software package (GraphPadSoftware, Inc., San Diego, Calif.) using the one-site competition model.K_(i) values for test compounds were calculated from observed IC₅₀values and the K_(D) value of the radioligand using the Cheng-Prusoffequation (Cheng Y; Prusoff W H. (1973) Biochemical Pharmacology,22(23):3099-108). K_(i) values were converted to pK_(i) values todetermine the geometric mean and 95% confidence intervals. These summarystatistics were then converted back to K_(i) values for data reporting.

In this assay, a lower K_(i) value indicates that the test compound hasa higher binding affinity for the receptor tested. Exemplified compoundof this invention that were tested in this assay typically were found tohave a K_(i) value of less than about 300 nM for the M₃ muscarinicreceptor. For example, the compounds of Examples 3 and 6 were found tohave K_(i) values of less than 10 nM.

Assay Test Procedure C

Whole-Cell Camp Flashplate Assay in CHO Cell Lines HeterologouslyExpressing Human β₁, β₂ or β₃ Adrenergic Receptors

cAMP assays were performed in a radioimmunoassay format using theFlashplate Adenylyl Cyclase Activation Assay System with [¹²⁵I]-cAMP(NEN SMP004, PerkinElmer Life Sciences Inc., Boston, Mass.), accordingto the manufacturers instructions. For the determination of β receptoragonist potency (EC₅₀), CHO-K1 cell lines stably expressing cloned humanβ₁, β₂ or β₃ adrenergic receptors were grown to near confluency in HAM'sF-12 media supplemented with 10% FBS and Geneticin (250 μg/mL). Cellswere rinsed with PBS and detached in dPBS (Dulbecco's Phosphate BufferedSaline, without CaCl₂ and MgCl₂) containing 2 mM EDTA or Trypsin-EDTAsolution (0.05% trypsin/0.53 mM EDTA). After counting cells in Coultercell counter, cells were pelleted by centrifugation at 1,000 rpm andre-suspended in stimulation buffer containing IBMX (PerkinElmer Kit)pre-warmed to room temperature to a concentration of 1.6×10⁶ to 2.8×10⁶cells/mL. About 60,000 to 80,000 cells per well were used in this assay.Test compounds (10 mM in DMSO) were diluted into PBS containing 0.1% BSAin Beckman Biomek-2000 and tested at 11 different concentrations rangingfrom 100 μM to 1 pM. Reactions were incubated for 10 min at 37° C. andstopped by adding 100 μL of cold detection buffer containing [¹²⁵I]-cAMP(NEN SMP004, PerkinElmer Life Sciences, Boston, Mass.). The amount ofcAMP produced (pmol/well) was calculated based on the counts observedfor the samples and cAMP standards as described in the manufacturer'suser manual. Data were analyzed by nonlinear regression analysis withthe GraphPad Prism Software package (GraphPad Software, Inc., San Diego,Calif.) with the sigmoidal equation. The Cheng-Prusoff equation (ChengY, and Prusoff W H., Biochemical Pharmacology, 1973, 22, 23, 3099-108)was used to calculate the EC50 values.

In this assay, a lower EC₅₀ value indicates that the test compound has ahigher functional activity at the receptor tested. Exemplified compoundof this invention that were tested in this assay typically were found tohave a EC₅₀ value of less than about 300 nM for the β₂ adrenergicreceptor. For example, the compounds of Examples 3 and 6 were found tohave EC₅₀ values of less than 10 nM.

If desired, the receptor subtype selectivity for a test compound can becalculated as the ratio of EC₅₀(β₁)/EC₅₀(β₂) or EC₅₀(β₃)/EC₅₀(β₂).Typically, compounds of this invention demonstrated greater functionalactivity at the β₂ adrenergic receptor compared to the β₁ or β₃adrenergic receptor, i.e. EC₅₀(β₁) or EC₅₀(β₃) is typically greater thanEC₅₀(β₂). Generally, compounds having selectivity for the β₂ adrenergicreceptor over the or β₃ adrenergic receptors are preferred; especiallycompounds having a selectivity greater than about 5; and in particular,greater than about 10. By way of example, the compounds of Examples 3and 6 had ratios of EC₅₀(β₁)/EC₅₀(β₂) greater than 10.

Assay Test Procedure D

Functional Assays of Antagonism for Muscarinic Receptor Subtypes

A. Blockade of Agonist-Mediated Inhibition of cAMP Accumulation

In this assay, the functional potency of a test compound is determinedby measuring the ability of the test compound to blockoxotremorine-inhibition of forskolin-mediated cAMP accumulation inCHO-K1 cells expressing the hM₂ receptor. cAMP assays are performed in aradioimmunoassay format using the Flashplate Adenylyl Cyclase ActivationAssay System with ¹²⁵I-cAMP (NEN SMP004B, PerkinElmer Life SciencesInc., Boston, Mass.), according to the manufacturer's instructions.Cells are rinsed once with dPBS and lifted with Trypsin-EDTA solution(0.05% trypsin/0.53 mM EDTA) as described in the Cell Culture andMembrane Preparation section above. The detached cells are washed twiceby centrifugation at 650×g for five minutes in 50 mL dPBS. The cellpellet is then re-suspended in 10 mL dPBS, and the cells are countedwith a Coulter Z1 Dual Particle Counter (Beckman Coulter, Fullerton,Calif.). The cells are centrifuged again at 650×g for five minutes andre-suspended in stimulation buffer to an assay concentration of1.6×10⁶-2.8×10⁶ cells/mL.

The test compound is initially dissolved to a concentration of 400 μM indilution buffer (dPBS supplemented with 1 mg/mL BSA (0.1%)), and thenserially diluted with dilution buffer to final molar concentrationsranging from 100 μM to 0.1 nM. Oxotremorine is diluted in a similarmanner.

To measure oxotremorine inhibition of adenylyl cyclase (AC) activity, 25μL forskolin (25 μM final concentration diluted in dPBS), 25 μL dilutedoxotremorine, and 50 μL cells are added to agonist assay wells. Tomeasure the ability of a test compound to block oxotremorine-inhibitedAC activity, 25 μL forskolin and oxotremorine (25 μM and 5 μM finalconcentrations, respectively, diluted in dPBS), 25 μL diluted testcompound, and 50 μL cells are added to remaining assay wells.

Reactions are incubated for 10 minutes at 37° C. and stopped by additionof 100 μL ice-cold detection buffer. Plates are sealed, incubatedovernight at room temperature and counted the next morning on aPerkinElmer TopCount liquid scintillation counter (PerkinElmer Inc.,Wellesley, Mass.). The amount of cAMP produced (pmol/well) is calculatedbased on the counts observed for the samples and cAMP standards, asdescribed in the manufacturer's user manual. Data is analyzed bynonlinear regression analysis with the GraphPad Prism Software package(GraphPad Software, Inc., San Diego, Calif.) using the non-linearregression, one-site competition equation. The Cheng-Prusoff equation isused to calculate the K_(i), using the EC₅₀ of the oxotremorineconcentration-response curve and the oxotremorine assay concentration asthe K_(D) and [L], respectively.

In this assay, a lower K_(i) value indicates that the test compound hasa higher functional activity at the receptor tested. Exemplifiedcompound of this invention that were tested in this assay typically werefound to have a K_(i) value of less than about 300 nM for blockade ofoxotremorine-inhibition of forskolin-mediated cAMP accumulation inCHO-K1 cells expressing the hM₂ receptor. For example, the compound ofExample 3 was found to have a K_(i) value of less than 10 nM.

B. Blockade of Agonist-Mediated [³⁵S]GTPγS Binding

In a second functional assay, the functional potency of test compoundscan be determined by measuring the ability of the compounds to blockoxotremorine-stimulated [³⁵S]GTPγS binding in CHO-K1 cells expressingthe hM₂ receptor.

At the time of use, frozen membranes were thawed and then diluted inassay buffer with a final target tissue concentration of 5-10 μg proteinper well. The membranes were briefly homogenized using a PolytronPT-2100 tissue disrupter and then added to the assay plates.

The EC₉₀ value (effective concentration for 90% maximal response) forstimulation of [³⁵S]GTPγS binding by the agonist oxotremorine wasdetermined in each experiment.

To determine the ability of a test compound to inhibitoxotremorine-stimulated [³⁵S]GTPγS binding, the following was added toeach well of 96 well plates: 25 μL of assay buffer with [³⁵S]GTPγS (0.4nM), 25 μL of oxotremorine (EC₉₀) and GDP (3 uM), 25 μL of diluted testcompound and 25 μL CHO cell membranes expressing the hM₂ receptor. Theassay plates were then incubated at 37° C. for 60 minutes. The assayplates were filtered over 1% BSA-pretreated GF/B filters using aPerkinElmer 96-well harvester. The plates were rinsed with ice-cold washbuffer for 3×3 seconds and then air or vacuum dried. Microscint-20scintillation liquid (50 μL) was added to each well, and each plate wassealed and radioactivity counted on a Topcounter (PerkinElmer). Datawere analyzed by nonlinear regression analysis with the GraphPad PrismSoftware package (GraphPad Software, Inc., San Diego, Calif.) using thenon-linear regression, one-site competition equation. The Cheng-Prusoffequation was used to calculate the K_(i), using the IC₅₀ values of theconcentration-response curve for the test compound and the oxotremorineconcentration in the assay as the K_(D) and [L], ligand concentration,respectively.

In this assay, a lower K_(i) value indicates that the test compound hasa higher functional activity at the receptor tested. Exemplifiedcompound of this invention that were tested in this assay typically werefound to have a K_(i) value of less than about 300 nM for blockade ofoxotremorine-stimulated [³⁵S]GTPγS binding in CHO-K1 cells expressingthe hM₂ receptor. For example, the compound of Example 3 was found tohave a K_(i) value of less than 10 nM.

C. Blockade of Agonist-Mediated Calcium Release via FLIPR Assays

Muscarinic receptor subtypes (M₁, M₃ and M₅ receptors), which couple toG_(q) proteins, activate the phospholipase C (PLC) pathway upon agonistbinding to the receptor. As a result, activated PLC hydrolyzesphosphatyl inositol diphosphate (PIP₂) to diacylglycerol (DAG) andphosphatidyl-1,4,5-triphosphate (IP₃), which in turn generates calciumrelease from intracellular stores, i.e., endoplasmic and sarcoplasmicreticulum. The FLIPR (Molecular Devices, Sunnyvale, Calif.) assaycapitalizes on this increase in intracellular calcium by using a calciumsensitive dye (Fluo-4AM, Molecular Probes, Eugene, Oreg.) thatfluoresces when free calcium binds. This fluorescence event is measuredin real time by the FLIPR, which detects the change in fluorescence froma monolayer of cells cloned with human M₁ and M₃, and chimpanzee M₅receptors. Antagonist potency can be determined by the ability ofantagonists to inhibit agonist-mediated increases in intracellularcalcium.

For FLIPR calcium stimulation assays, CHO cells stably expressing thehM₁, hM₃ and cM₅ receptors are seeded into 96-well FLIPR plates thenight before the assay is done. Seeded cells are washed twice byCellwash (MTX Labsystems, Inc.) with FLIPR buffer (10 mM HEPES, pH 7.4,2 mM calcium chloride, 2.5 mM probenecid in Hank's Buffered SaltSolution (HBSS) without calcium and magnesium) to remove growth mediaand leaving 50 μL/well of FLIPR buffer. The cells are then incubatedwith 50 μL/well of 4 μM FLUO-4AM (a 2× solution was made) for 40 minutesat 37° C., 5% carbon dioxide. Following the dye incubation period, cellsare washed two times with FLIPR buffer, leaving a final volume of 50μL/well.

To determine antagonist potency, the dose-dependent stimulation ofintracellular Ca²⁺ release for oxotremorine is first determined so thatantagonist potency can later be measured against oxotremorinestimulation at an EC₉₀ concentration. Cells are first incubated withcompound dilution buffer for 20 minutes, followed by agonist addition,which is performed by the FLIPR. An EC₉₀ value for oxotremorine isgenerated according to the method detailed in the FLIPR measurement anddata reduction section below, in conjunction with the formulaEC_(F)=((F/100−F)ˆ1/H)*EC₅₀. An oxotremorine concentration of 3×EC_(F)is prepared in stimulation plates such that an EC₉₀ concentration ofoxotremorine is added to each well in the antagonist inhibition assayplates.

The parameters used for the FLIPR are: exposure length of 0.4 seconds,laser strength of 0.5 watts, excitation wavelength of 488 nm, andemission wavelength of 550 nm. Baseline is determined by measuring thechange in fluorescence for 10 seconds prior to addition of agonist.Following agonist stimulation, the FLIPR continuously measured thechange of fluorescence every 0.5 to 1 second for 1.5 minutes to capturethe maximum fluorescence change.

The change of fluorescence is expressed as maximum fluorescence minusbaseline fluorescence for each well. The raw data is analyzed againstthe logarithm of drug concentration by nonlinear regression withGraphPad Prism (GraphPad Software, Inc., San Diego, Calif.) using thebuilt-in model for sigmoidal dose-response. Antagonist K_(i) values aredetermined by Prism using the oxotremorine EC₅₀ value as the K_(D) andthe oxotremorine EC₉₀ for the ligand concentration according to theCheng-Prusoff equation (Cheng & Prusoff, 1973).

In this assay, a lower K_(i) value indicates that the test compound hasa higher functional activity at the receptor tested. Exemplifiedcompound of this invention that were tested in this assay typically werefound to have a K_(i) value of less than about 300 nM for blockade ofagonist-mediated calcium release in CHO cells stably expressing the hM₁,hM₃ and cM₅ receptors. For example, the compound of Example 3 was foundto have a K value of less than 10 nM for the hM₁, hM₃ and cM₅ receptors.

Assay Test Procedure E

Whole-Cell Camp Flashplate Assay with a Lung Epithelial Cell LineEndogenously Expressing Human β₂ Adrenergic Receptor

For the determination of agonist potencies and efficacies (intrinsicactivities) in a cell line expressing endogenous levels of the β₂adrenergic receptor, a human lung epithelial cell line (BEAS-2B) wasused (ATCC CRL-9609, American Type Culture Collection, Manassas, Va.)(January B, et al., British Journal of Pharmacology, 1998, 123, 4,701-11). Cells were grown to 75-90% confluency in complete, serum-freemedium (LHC-9 MEDIUM containing Epinephrine and Retinoic Acid, cat#181-500, Biosource International, Camarillo, Calif.). The day beforethe assay, medium was switched to LHC-8 (no epinephrine or retinoicacid, cat #141-500, Biosource International, Camarillo, Calif.). cAMPassays were performed in a radioimmunoassay format using the FlashplateAdenylyl Cyclase Activation Assay System with [¹²⁵I]-cAMP (NEN SMP004,PerkinElmer Life Sciences Inc., Boston, Mass.), according to themanufacturers instructions. On the day of the assay, cells were rinsedwith PBS, lifted by scraping with 5 mM EDTA in PBS, and counted. Cellswere pelleted by centrifugation at 1,000 rpm and re-suspended instimulation buffer pre-warmed to 37° C. at a final concentration of600,000 cells/mL. Cells were used at a final concentration of 100,000 to120,000 cells/well in this assay. Test compounds were serially dilutedinto assay buffer (75 mM Tris/HCl pH 7.4 at 25° C., 12.5 mM MgCl₂, 1 mMEDTA, 0.2% BSA) in Beckman Biomek-2000. Test compounds were tested inthe assay at 11 different concentrations, ranging from 10 μM to 10 pM.Reactions were incubated for 10 min at 37° C. and stopped by addition of100 μL of ice-cold detection buffer. Plates were sealed, incubated overnight at 4° C. and counted the next morning in a Topcount scintillationcounter (Packard BioScience Co., Meriden, Conn.). The amount of cAMPproduced per mL of reaction was calculated based on the counts observedfor samples and cAMP standards, as described in the manufacturer's usermanual. Data were analyzed by nonlinear regression analysis with theGraphPad Prism Software package (GraphPad Software, Inc., San Diego,Calif.) using the 4-parameter model for sigmoidal dose-response.

In this assay, a lower EC₅₀ value indicates that the test compound has ahigher functional activity at the receptor tested. Exemplified compoundof this invention that were tested in this assay typically were found tohave a EC₅₀ value of less than about 300 nM for the β₂ adrenergicreceptor. For example, the compounds of Examples 3 and 6 were found tohave EC₅₀ values of less than 10 nM.

If desired, test compound efficacy (% Eff) was calculated from the ratioof the observed Emax (TOP of the fitted curve) and the maximal responseobtained for isoproterenol dose response curve and was expressed as %Eff relative to isoproterenol. Exemplified compounds of this inventiontested in this assay typically demonstrated a % Eff greater than about40.

Assay Test Procedure F

Duration of Bronchoprotection in Guinea Pig Models ofAcetylcholine-Induced or Histamine-Induced Bronchoconstriction

These in vivo assays were used to assess the bronchoprotective effectsof test compounds exhibiting both muscarinic receptor antagonist and β₂adrenergic receptor agonist activity. To isolate muscarinic antagonistactivity in the acetylcholine-induced bronchoconstriction model, theanimals were administered propanolol, a compound that blocks β receptoractivity, prior to the administration of acetylcholine. Duration ofbronchoprotection in the histamine-induced bronchoconstriction modelreflects β₂ adrenergic receptor agonist activity.

Groups of 6 male guinea pigs (Duncan-Hartley (HsdPoc:DH) Harlan,Madison, Wis.) weighing between 250 and 350 g were individuallyidentified by cage cards. Throughout the study, animals were allowedaccess to food and water ad libitum.

Test compounds were administered via inhalation over 10 minutes in awhole-body exposure dosing chamber (R&S Molds, San Carlos, Calif.). Thedosing chambers were arranged so that an aerosol was simultaneouslydelivered to 6 individual chambers from a central manifold. Guinea pigswere exposed to an aerosol of a test compound or vehicle (WFI). Theseaerosols were generated from aqueous solutions using an LC StarNebulizer Set (Model 22F51, PARI Respiratory Equipment, Inc. Midlothian,Va.) driven by a mixture of gases (CO₂=5%, O₂=21% and N₂=74%) at apressure of 22 psi. The gas flow through the nebulizer at this operatingpressure was approximately 3 L/minute. The generated aerosols weredriven into the chambers by positive pressure. No dilution air was usedduring the delivery of aerosolized solutions. During the 10 minutenebulization, approximately 1.8 mL of solution was nebulized. This valuewas measured gravimetrically by comparing pre- and post-nebulizationweights of the filled nebulizer.

The bronchoprotective effects of test compounds administered viainhalation were evaluated using whole body plethysmography at 1.5, 24,48 and 72 hours post-dose.

Forty-five minutes prior to the start of the pulmonary evaluation, eachguinea pig was anesthetized with an intramuscular injection of ketamine(43.75 mg/kg), xylazine (3.50 mg/kg) and acepromazine (1.05 mg/kg).After the surgical site was shaved and cleaned with 70% alcohol, a 2-3cm midline incision of the ventral aspect of the neck was made. Then,the jugular vein was isolated and cannulated with a saline-filledpolyethylene catheter (PE-50, Becton Dickinson, Sparks, Md.) to allowfor intravenous infusions of acetylcholine (Ach) or histamine in saline.The trachea was then dissected free and cannulated with a 14G teflontube (#NE-014, Small Parts, Miami Lakes, Fla.). If required, anesthesiawas maintained by additional intramuscular injections of theaforementioned anesthetic mixture. The depth of anesthesia was monitoredand adjusted if the animal responds to pinching of its paw or if therespiration rate was greater than 100 breaths/minute.

Once the cannulations were completed, the animal was placed into aplethysmograph (#PLY3114, Buxco Electronics, Inc., Sharon, Conn.) and anesophageal pressure cannula (PE-160, Becton Dickinson, Sparks, Md.) wasinserted to measure pulmonary driving pressure (pressure). The teflontracheal tube was attached to the opening of the plethysmograph to allowthe guinea pig to breathe room air from outside the chamber. The chamberwas then sealed. A heating lamp was used to maintain body temperatureand the guinea pig's lungs were inflated 3 times with 4 mL of air usinga 10 mL calibration syringe (#5520 Series, Hans Rudolph, Kansas City,Mo.) to ensure that the lower airways did not collapse and that theanimal did not suffer from hyperventilation.

Once it was determined that baseline values were within the range of0.3-0.9 mL/cm H₂O for compliance and within the range of 0.1-0.199 cmH₂O/mL per second for resistance, the pulmonary evaluation wasinitiated. A Buxco pulmonary measurement computer program enabled thecollection and derivation of pulmonary values.

Starting this program initiated the experimental protocol and datacollection. The changes in volume over time that occur within theplethysmograph with each breath were measured via a Buxco pressuretransducer. By integrating this signal over time, a measurement of flowwas calculated for each breath. This signal, together with the pulmonarydriving pressure changes, which were collected using a Sensym pressuretransducer (#TRD4100), was connected via a Buxco (MAX 2270) preamplifierto a data collection interface (#'s SFT3400 and SFT3813). All otherpulmonary parameters were derived from these two inputs.

Baseline values were collected for 5 minutes, after which time theguinea pigs were challenged with Ach or histamine. When evaluating themuscarinic antagonist effects, propanolol (5 mg/Kg, iv) (Sigma-Aldrich,St. Louis, Mo.) was administered 15 minutes prior to challenge with Ach.Ach (Sigma-Aldrich, St. Louis, Mo.) (0.1 mg/mL) was infusedintravenously for 1 minute from a syringe pump (sP210iw, World PrecisionInstruments, Inc., Sarasota, Fla.) at the following doses and prescribedtimes from the start of the experiment: 1.9 μg/minute at 5 minutes, 3.8μg/minute at 10 minutes, 7.5 μg/minute at 15 minutes, 15.0 μg/minute at20 minutes, 30 μg/minute at 25 minutes and 60 μg/minute at 30 minutes.Alternatively, bronchoprotection of test compounds was assessed in theacetylcholine challenge model without pretreatment with a beta blockingcompound.

When evaluating the β₂ adrenergic receptor agonist effects of testcompounds, histamine (25 μg/mL) (Sigma-Aldrich, St. Louis, Mo.) wasinfused intravenously for 1 minute from a syringe pump at the followingdoses and prescribed times from the start of the experiment: 0.5μg/minute at 5 minutes, 0.9 μg/minute at 10 minutes, 1.9 μg/minute at 15minutes, 3.8 μg/minute at 20 minutes, 7.5 μg/minute at 25 minutes and 15μg/minute at 30 minutes. If resistance or compliance had not returned tobaseline values at 3 minutes following each Ach or histamine dose, theguinea pig's lungs were inflated 3 times with 4 mL of air from a 10 mLcalibration syringe. Recorded pulmonary parameters include respirationfrequency (breaths/minute), compliance (mL/cm H₂O) and pulmonaryresistance (cm H₂O/mL per second). Once the pulmonary functionmeasurements were completed at minute 35 of this protocol, the guineapig was removed from the plethysmograph and euthanized by carbon dioxideasphyxiation.

The data were evaluated in one of two ways:

(a) Pulmonary resistance (R_(L), cm H₂O/mL per second) was calculatedfrom the ratio of “change in pressure” to “the change in flow.” TheR_(L) response to ACh (60 μg/min, 1H) was computed for the vehicle andthe test compound groups. The mean ACh response in vehicle-treatedanimals, at each pre-treatment time, was calculated and used to compute% inhibition of ACh response, at the corresponding pre-treatment time,at each test compound dose. Inhibition dose-response curves for ‘R_(L)’were fitted with a four parameter logistic equation using GraphPadPrism, version 3.00 for Windows (GraphPad Software, San Diego, Calif.)to estimate bronchoprotective ID₅₀ (dose required to inhibit the ACh (60μg/min) bronchocontrictor response by 50%). The equation used was asfollows:Y=Min+(Max−Min)/(1+10^(((log ID50−X)*Hillslope)))where X is the logarithm of dose, Y is the response (% Inhibition of AChinduced increase in R_(L))— Y starts at Min and approachesasymptotically to Max with a sigmoidal shape.

(b) The quantity PD₂, which is defined as the amount of Ach or histamineneeded to cause a doubling of the baseline pulmonary resistance, wascalculated using the pulmonary resistance values derived from the flowand the pressure over a range of Ach or histamine challenges using thefollowing equation (derived from the equation used to calculate PC₂₀values in the clinic (see Am. Thoracic Soc, 2000):${PD}_{2} = {{antilog}\quad\left\lbrack {{\log\quad C_{1}} + \frac{\left( {{\log\quad C_{2}} - {\log\quad C_{1}}} \right)\left( {{2R_{0}} - R_{1}} \right)}{R_{2} - R_{1}}} \right\rbrack}$where:

-   -   C₁=concentration of Ach or histamine preceding C₂    -   C₂=concentration of Ach or histamine resulting in at least a        2-fold increase in pulmonary resistance (R_(L))    -   R₀=Baseline R_(L) value    -   R₁═R_(L) value after C₁    -   R₂═R_(L) value after C₂

Statistical analysis of the data was performed using a twotailed—Students t-test. A P-value <0.05 was considered significant.

Exemplified compounds of this invention that were tested in this assaytypically produced a dose-dependent bronchoprotective effect againstMCh-induced bronchoconstriction and His-induced bronchoconstriction.Generally, test compounds having a potency (ID₅₀ at 1.5 h post-dose) ofless than about 300 μg/mL for ACh-induced bronchoconstriction and lessthan about 300 μg/mL for His-induced bronchoconstriction in this assayare generally preferred. For example, the compounds of Examples 3 and 6were found to have an ID₅₀ less than about 100 μg/mL for ACh-inducedbronchoconstriction and an ID₅₀ less than about 100 μg/mL forHis-induced bronchoconstriction at 1.5 hours post-dose.

Additionally, test compounds having a duration (PD T_(1/2)) ofbrochoprotective activity of at least about 24 hours in this assay aregenerally preferred. By way of example, the compounds of Examples 3 and6 were found to have a PD T_(1/2) of at least about 24 hours post-dose.

Assay Test Procedure G

Einthoven Model for Measuring Changes in Ventilation in Guinea Pigs

The bronchodilator activity of test compounds was evaluated in ananesthetized guinea pig model (the Einthoven model), which usesventilation pressure as a surrogate measure of airway resistance. See,for example, Einthoven (1892) Pfugers Arch. 51: 367-445; and Mohammed etal. (2000) Pulm Pharmacol Ther. 13(6):287-92. In this model, muscarinicantagonist and β₂ agonist activity was assessed by determining theprotective effects against methacholine (MCh) and histamine(His)-induced bronchoconstriction.

This assay was conducted using Duncan-Hartley guinea pigs (Harlan,Indianapolis, Ind.), weighing between 300 and 400 g.

The test compound or vehicle (i.e., sterile water) was dosed byinhalation (1H) over a 10 minute time period in a whole body exposuredosing chamber (R+S Molds, San Carlos, Calif.) using 5 mL of dosingsolution. Animals were exposed to an aerosol, which was generated froman LC Star Nebulizer Set (Model 22F51, PARI Respiratory Equipment, Inc.Midlothian, Va.) driven by Bioblend a mixture of gasses (5% CO₂; 21% O₂;and 74% N₂) at a pressure of 22 psi. Pulmonary function was evaluated atvarious time-points after inhalation dosing.

Forty five minutes prior to the start of pulmonary function evaluation,the guinea pigs were anesthetized with an intramuscular (IM) injectionof a mixture of ketamine (13.7 mg/kg/xylazine (3.5 mg/kg)/acepromazine(1.05 mg/kg). A supplemental dose of this mixture (50% of initial dose)was administered as needed. The jugular vein and carotid artery wereisolated and cannulated with saline-filled polyethylene catheters(micro-renathane and PE-50, respectively, Beckton Dickinson, Sparks,Md.). The carotid artery was connected to a pressure transducer to allowthe measurement of blood pressure and the jugular vein cannula was usedfor IV injection of either MCh or His. The trachea was then dissectedfree and cannulated with a 14G needle (#NE-014, Small Parts, MiamiLakes, Fla.). Once the cannulations were complete, the guinea pigs wereventilated using a respirator (Model 683, Harvard Apparatus, Inc., MA)set at a stroke volume of 1 mL/100 g body weight but not exceeding 2.5mL volume, and at a rate of 100 strokes per minute. Ventilation pressure(VP) was measured in the tracheal cannula using a Biopac transducer thatwas connected to a Biopac (TSD 137C) pre-amplifier. Body temperature wasmaintained at 37° C. using a heating pad. Prior to initiating datacollection, pentobarbital (25 mg/kg) was administered intraperitoneally(IP) to suppress spontaneous breathing and obtain a stable baseline. Thechanges in VP were recorded on a Biopac Windows data collectioninterface. Baseline values were collected for at least 5 minutes, afterwhich time guinea pigs were challenged IV non-cumulatively with 2-foldincremental doses of the bronchoconstrictor (MCh or His). When MCh wasused as the bronchoconstrictor agent, animals were pre-treated withpropranolol (5 mg/kg, IV) to isolate the antimuscarinic effects of thetest compound. Changes in VP were recorded using the Acknowledge DataCollection Software (Santa Barbara, Calif.). After the completion ofstudy, the animals were euthanized.

Change in VP was measured in cm of water. Change in VP (cm H₂O)=peakpressure (after bronchoconstrictor challenge)−peak baseline pressure.The dose-response curve to MCh or His was fitted to a four parameterlogistic equation using GraphPad Prism, version 3.00 for Windows(GraphPad Software, San Diego, Calif.) The equation used was as follows:Y=Min+(Max−Min)/(1+10^(((log ID50−X)*Hilislope)))where X is the logarithm of dose, Y is the response. Y starts at Min andapproaches asymptotically to Max with a sigmoidal shape.

The percent inhibition of the bronchoconstrictor response to asubmaximal dose of MCh or His was calculated at each dose of the testcompound using the following equation: % Inhibition ofresponse=100−((peak pressure (after bronchoconstrictor challenge,treated)−peak baseline pressure (treated)*100%/(peak pressure (afterbronchoconstrictor challenge, water)−peak baseline pressure (water)).Inhibition curves were fitted using the four parameter logistic equationfrom GraphPad software. ID₅₀ (dose required to produce 50% inhibition ofthe bronchoconstrictor response) and Emax (maximal inhibition) were alsoestimated wherever appropriate.

The magnitude of bronchoprotection at different time-points afterinhalation of the test compound was used to estimate the pharmacodynamichalf-life (PD T_(1/2)). PD T_(1/2) was determined using a non-linearregression fit using a one-phase exponential decay equation (GraphPadPrism, Version 4.00): Y=Span*exp(−K*X)+Plateau; Starts at Span+Plateauand decays to Plateau with a rate constant K. The PD T_(1/2)=0.69/K.Plateau was constrained to 0.

Exemplified compounds of this invention that were tested in this assaytypically produced a dose-dependent bronchoprotective effect againstMCh-induced bronchoconstriction and His-induced bronchoconstriction.Generally, test compounds having an ID₅₀ less than about 300 μg/mL forMCh-induced bronchoconstriction and an ID₅₀ less than about 300 μg/mLfor His-induced bronchoconstriction at 1.5 hours post-dose in this assayare preferred. Additionally, test compounds having a duration (PDT_(1/2)) of brochoprotective activity of at least about 24 hours in thisassay are generally preferred.

Assay Test Procedure H

Inhalation Guinea Pig Salivation Assay

Guinea pigs (Charles River, Wilmington, Mass.) weighing 200-350 g wereacclimated to the in-house guinea pig colony for at least 3 daysfollowing arrival. Test compound or vehicle were dosed via inhalation(1H) over a 10 minute time period in a pie shaped dosing chamber (R+SMolds, San Carlos, Calif.). Test solutions were dissolved in sterilewater and delivered using a nebulizer filled with 5.0 mL of dosingsolution. Guinea pigs were restrained in the inhalation chamber for 30minutes. During this time, guinea pigs were restricted to an area ofapproximately 110 sq. cm. This space was adequate for the animals toturn freely, reposition themselves, and allow for grooming. Following 20minutes of acclimation, guinea pigs were exposed to an aerosol generatedfrom a LS Star Nebulizer Set (Model 22F51, PARI Respiratory Equipment,Inc. Midlothian, Va.) driven by house air at a pressure of 22 psi. Uponcompletion of nebulization, guinea pigs were evaluated at 1.5, 6, 12,24, 48, or 72 hrs after treatment.

Guinea pigs were anesthetized one hour before testing with anintramuscular (IM) injection of a mixture of ketamine 43.75 mg/kg,xylazine 3.5 mg/kg, and acepromazine 1.05 mg/kg at an 0.88 mL/kg volume.Animals were placed ventral side up on a heated (37° C.) blanket at a 20degree incline with their head in a downward slope. A 4-ply 2×2 inchgauze pad (Nu-Gauze General-use sponges, Johnson and Johnson, Arlington,Tex.) was inserted in the guinea pig's mouth. Five minutes later, themuscarinic agonist pilocarpine (3.0 mg/kg, s.c.) was administered andthe gauze pad was immediately discarded and replaced by a newpre-weighed gauze pad. Saliva was collected for 10 minutes, at whichpoint the gauze pad was weighed and the difference in weight recorded todetermine the amount of accumulated saliva (in mg). The mean amount ofsaliva collected for animals receiving the vehicle and each dose of testcompound was calculated. The vehicle group mean was considered to be100% salivation. Results were calculated using result means (n=3 orgreater). Confidence intervals (95%) were calculated for each dose ateach time point using two-way ANOVA. This model is a modified version ofthe procedure described in Rechter, “Estimation of anticholinergic drugeffects in mice by antagonism against pilocarpine-induced salivation”Ata Pharmacol Toxicol, 1996, 24:243-254.

The mean weight of saliva in vehicle-treated animals, at eachpre-treatment time, was calculated and used to compute % inhibition ofsalivation, at the corresponding pre-treatment time, at each dose. Theinhibition dose-response data were fitted to a four parameter logisticequation using GraphPad Prism, version 3.00 for Windows (GraphPadSoftware, San Diego, Calif.) to estimate anti-sialagogue ID₅₀ (doserequired to inhibit 50% of pilocarpine-evoked salivation). The equationused was as follows:Y=Min+(Max−Min)/(1+10^(((log ID50−X)*Hilislope)))where X is the logarithm of dose, Y is the response (% inhibition ofsalivation). Y starts at Min and approaches asymptotically to Max with asigmoidal shape.

The ratio of the anti-sialagogue ID₅₀ to bronchoprotective ID₅₀ was usedto compute the apparent lung-selectivity index of the test compound.Generally, compounds having an apparent lung-selectivity index greaterthan about 5 are preferred. In this assay, the compound of Example 3 hadan apparent lung-selectivity index greater than 5.

While the present invention has been described with reference tospecific aspects or embodiments thereof, it will be understood by thoseof ordinary skilled in the art that various changes can be made orequivalents can be substituted without departing from the true spiritand scope of the invention. Additionally, to the extent permitted byapplicable patent statues and regulations, all publications, patents andpatent applications cited herein are hereby incorporated by reference intheir entirety to the same extent as if each document had beenindividually incorporated by reference herein.

1-43. (canceled)
 44. A pharmaceutical composition comprising apharmaceutically acceptable carrier and a therapeutically effectiveamount of a compound of the formula:

wherein o is an integer from 2 to 7; p is an integer from 1 to 6;provided that o+p is an integer from 3 to 8; the phen-1,4-ylene group isoptionally substituted with from 1 to 4 substituents independentlyselected from halo, (1-4C)alkyl, (1-4C)alkoxy, —S-(1-4C)alkyl,—S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl, —C(O)O(1-4C)alkyl, carboxy,cyano, hydroxy, nitro, trifluoromethyl and trifluoromethoxy; or apharmaceutically acceptable salt or stereoisomer thereof.
 45. Thepharmaceutical composition of claim 44, wherein o is 2 or
 3. 46. Thepharmaceutical composition of claim 44, wherein o is
 2. 47. Thepharmaceutical composition of claim 44, wherein p is 1 or
 2. 48. Thepharmaceutical composition of claim 44, wherein p is
 1. 49. Thepharmaceutical composition of claim 44, wherein o is 2 and p is
 1. 50.The pharmaceutical composition of claim 44, wherein the phen-1,4-ylenegroup has one substituent.
 51. The pharmaceutical composition of claim44, wherein the phen-1,4-ylene group has two substituents.
 52. Thepharmaceutical composition of claim 44, wherein the phen-1,4-ylene groupis substituted with from 1 to 4 substituents and at least one of thesubstituents is halo.
 53. The pharmaceutical composition of claim 44,wherein the phen-1,4-ylene group is substituted with from 1 to 4substituents and at least one of the substituents is chloro.
 54. Thepharmaceutical composition of claim 44, wherein the phen-1,4-ylene groupis substituted with from 1 to 4 substituents and at least one of thesubstituents is (1-4C)alkoxy.
 55. The pharmaceutical composition ofclaim 44, wherein the phen-1,4-ylene group is substituted with from 1 to4 substituents and at least one of the substituents is methoxy.
 56. Thepharmaceutical composition of claim 44, wherein the phen-1,4-ylene groupis substituted with from 1 to 4 substituents and at least one of thesubstituents is (1-4C)alkyl.
 57. The pharmaceutical composition of claim44, wherein the phen-1,4-ylene group is substituted with from 1 to 4substituents and at least one of the substituents is methyl.
 58. Thepharmaceutical composition of claim 44, wherein the compound has the (R)configuration at the carbon atom of the group —CH(OH)—.
 59. Thepharmaceutical composition of claim 44, wherein the compound is in theform of a pharmaceutically acceptable salt.
 60. The pharmaceuticalcomposition of claim 44, wherein the compound is in the form of a freebase.
 61. The pharmaceutical composition of claim 44, wherein thecomposition is an aerosol or a powder.
 62. The pharmaceuticalcomposition of claim 44, wherein the composition is micronized.
 63. Amethod of treating a patient having chronic obstructive pulmonarydisease or asthma, the method comprising administering to the patient atherapeutically effective amount of a composition of any one of claims44 to
 62. 64. A method of producing bronchodilation in a patient, themethod comprising administering to the patient a therapeuticallyeffective amount of a composition of any one of claims 44 to 62.